Isolation of RNA from milk somatic cells as an alternative to biopsies of mammary tissue for nutrigenomic studies in dairy ewes

11 páginas, 4 tablas, 1 figura.Nutrigenomic studies of mammary lipogenesis in ruminants often rely on the use of mammary tissue (MT) collected either by biopsy or at slaughter. However, isolating RNA from milk would be a useful and cost-effective technique that may avoid distress to the animal and facilitate the collection of samples in time series experiments. This assay was therefore conducted to test the hypothesis that RNA extracted from milk somatic cells (MSC) in dairy sheep would be a feasible alternative to the performance of MT biopsies for nutrigenomic analyses. To meet this objective, 8 lactating Assaf ewes were divided in 2 groups and offered a total mixed ration without supplementation (control) or supplemented with 2.4% dry matter of fish oil, which was known not only to elicit milk fat depression but also to downregulate the expression of some candidate genes involved in mammary lipogenesis. Total RNA was extracted from MSC and biopsied MT to examine whether the potential changes in the abundance of transcripts was similarly detected with both RNA sources. Milk fatty acid profile was also analyzed by gas chromatography, and variations in mRNA abundance were determined by reverse transcription quantitative PCR. Values of RNA integrity number were always ≥7.7. The expected and designed decrease of milk fat concentration with fish oil (−29%), was associated with a lower transcript abundance of genes coding for enzymes involved in fatty acid activation (ACSS1), de novo synthesis (ACACA and FASN), uptake from plasma lipids (LPL), and esterification of fatty acids to glycerol (LPIN1), as well as of a transcription factor that may regulate their expression (INSIG1). Stable mRNA levels were showed in other candidate genes, such as FABP3, GPAT4, or SCD. Changes due to the dietary treatment were similarly detected with both RNA sources (MSC and MT biopsies), which supports the initial hypothesis and would validate the use of milk as an alternative RNA source for nutrigenomic analyses in dairy sheep.This work was supported by the Ministry of Economy and Competitiveness (MINECO, Spain; AGL2014-54587, co-funded by the European Regional Development Fund, European Union). P. G. Toral gratefully acknowledges receipt of a postdoctoral research contract from the MINECO (Juan de la Cierva program).Peer reviewe

Heparan sulfate proteoglycans undergo differential expression alterations in left sided colorectal cancer, depending on their metastatic character

Abstract Background Heparan sulfate proteoglycans (HSPGs) are complex molecules which play a role in the invasion and growth and metastatic properties of cancerous cells. In this work we analyze changes in the patterns of expression of HSPGs in left sided colorectal cancer (LSCRC), both metastatic and non-metastatic, and the results are also compared with those previously obtained for right sided tumors (RSCRCs). Methods Eighteen LSCRCs were studied using qPCR to analyze the expression of both the proteoglycan core proteins and the enzymes involved in heparan sulfate chain biosynthesis. Certain HSPGs also carry chondroitin sulfate chains and so we also studied the genes involved in its biosynthesis. The expression of certain genes that showed significant expression differences were also analysed using immunohistochemical techniques. Results Changes in proteoglycan core proteins were dependent on their location, and the main differences between metastatic and non-metastatic tumors affected cell-surface glypicans, while other molecules were quite similar. Glypicans were also responsible for the main differences between RS- and LS- malignances. Regarding the biosynthesis of heparan sulfate chains, differential alterations in transcription depending on the presence or not of metastasis affected genes involved in the modification of uronic acid (epimerization and 2-O sulfation), and some isoforms responsible for sulfation of glucosamine (NDST1, HS6ST1). Moreover, in RSCRCs differences were preferentially found in the expression of genes involved in C6 and C3 sulfation of glucosamine, but not in NDSTs or SULFs. Finally, synthesis of chondroitin sulfate showed some alterations, which affected various steps, including polimerization and the modification of chains, but the main variations dependent on the presence of metastases were epimerization and 6C sulfation; however, when compared with RSCRCs, the essential divergences affected polymerization of the chains and the 6C sulfation of the galactosamine residue. Conclusions We evidenced alterations in the expression of HSPGs, including the expression of cell surface core proteins, many glycosiltransferases and some enzymes that modify the GAG chains in LSCRCs, but this was dependent on the metastatic nature of the tumor. Some of these alterations are shared with RSCRCs, while others, focused on specific gene groups, are dependent on tumor localization

Elucidating fish oil-induced milk fat depression in dairy sheep: Milk somatic cell transcriptome analysis

9 páginas, 4 figuras.In this study, RNA sequencing was used to obtain a comprehensive profile of the transcriptomic changes occurring in the mammary gland of lactating sheep suffering from fish oil-induced milk fat depression (FO-MFD). The milk somatic cell transcriptome analysis of four control and four FO-MFD ewes generated an average of 42 million paired-end reads per sample. In both conditions, less than 220 genes constitute approximately 89% of the total counts. These genes, which are considered as core genes, were mainly involved in cytoplasmic ribosomal proteins and electron transport chain pathways. In total, 117 genes were upregulated, and 96 genes were downregulated in FO-MFD samples. Functional analysis of the latter indicated a downregulation of genes involved in the SREBP signaling pathway (e.g., ACACA, ACSL, and ACSS) and Gene Ontology terms related to lipid metabolism and lipid biosynthetic processes. Integrated interpretation of upregulated genes indicated enrichment in genes encoding plasma membrane proteins and proteins regulating protein kinase activity. Overall, our results indicate that FO-MFD is associated with the downregulation of key genes involved in the mammary lipogenesis process. In addition, the results also suggest that this syndrome may be related to upregulation of other genes implicated in signal transduction and codification of transcription factors.This work was supported by the Spanish Ministry of Economy and Competitiveness (MINECO; AGL2014-54587-R and AGL2015-66035-R projects, co-funded by the European Regional Development Fund). P.G.T. and B.G.G. are funded by two postdoctoral research contracts from MINECO (Juan de la Cierva -JCI-2012-11836- and Ramón y Cajal -RYC-2012-10230 programs, respectively).Peer reviewe

Estudo de graduados da Faculdade de Ciências da Saúde, coortes 2014 e 2015, Boyacá, Colômbia

Introduction: The graduate studies evaluate professional practice, in which characteristics such as profile, acquired competences, employment status, identity with the institution of higher education and the perception of teacher training are described, allowing establishing improvement actions. Objective: Describe sociodemographic characteristics, employment situation, satisfaction and identity perceived by the graduates of the programs of the Faculty of Health Sciences at the time of the degree in the 2014 and 2015 cohorts.Materials y methods: A descriptive study with a single cohort design, stratified sampling, and a sample of 120 graduates was carried out.Results: The average age of completion of secondary education was 16 years; the academic formation of the father and the mother evidenced the completion of secondary education, with the mothers being the highest percentage culminating university studies.After completing high school, 43% of graduates entered the university in less than 3 months. Twenty-two per cent work at IPS. A high percentage expressed that they would return to carry out undergraduate studies in the institution attending to the quality of the offered training, followed by the quality of professors; compared to investigative processes refer 16.7% to have participated in a research group. Regarding the competences acquired by the graduates, they show satisfaction with the communicative skills, leadership, coping with change, research and information processing. Conclusions: The training imparted by the university was perceived as directly related to the job performance, being satisfied with the communicative, investigative and information processing skills.Introducción. Los estudios de graduados evalúan el ejercicio profesional y en ellos se describen características como el perfil, las competencias adquiridas, la situación laboral, la identidad con la institución de educación superior y la percepción frente a la formación docente, permitiendo establecer acciones de mejoramiento. Objetivo. Describir las características sociodemográficas, la situación laboral, la satisfacción y la identidad percibida por los egresados de los programas de la Facultad de Ciencias de la Salud al momento del grado, en las cohortes 2014 y 2015. Materiales y métodos. Se llevó a cabo un estudio descriptivo con diseño de cohorte única, muestreo estratificado y muestra de 120 graduados. Resultados. La edad promedio de finalización de los estudios de secundaria fue de 16 años; la formación académica del padre y de la madre evidenció la finalización de la secundaria completa, y las madres eran quienes en mayor porcentaje habían culminado sus estudios universitarios. Después de culminar el bachillerato, el 43 % de los graduados ingresaron en menos de tres meses a la universidad. El 22 % trabaja en instituciones prestadoras de salud (IPS). El 39.2% expresó que volvería a adelantar estudios de pregrado en la institución atendiendo a la calidad de la formación ofrecida, seguida por la calidad de losIntrodução. Os estudos de graduados avaliam a prática profissional e descrevem características como perfil, habilidades adquiridas, situação laboral, identidade com a instituição de ensino superior e percepção de formação de professores, permitindo ações de melhora.Objetivo. Descrever as características sócio-demográficas, a situação do emprego, a satisfação e a identidade percebidas pelos graduados dos programas da Faculdade de Ciências da Saúde no momento de terminar a graduação, nas coortes 2014 e 2015.Materiais e métodos. Foi realizado um estudo descritivo com um único coorte, amostragem estratificada e amostra de 120 graduados.Resultados. A idade média de conclusão do ensino secundário foi de 16 anos; a formação acadêmica do pai e a mãe evidenciaram a conclusão do ensino médio completo, e as mães apresentaram maior porcentagem de estudos universitários finalizados.Depois de completar o ensino médio, 43% dos participantes entraram na universidade em menos de três meses. 22% trabalham em instituições de saúde. 39,2% manifestaram que retornariam para realizar estudos de graduação na instituição atendendo à qualidade da formação oferecida, seguido pela qualidade dos professores; em relação aos processos de pesquisa, 16,7% relataram ter participado de um grupo de pesquisa. Em relação às competências adquiridas pelos graduados, eles disseram estar satisfeitos com habilidades de comunicação, liderança, enfrentamento às mudanças e processamento das informações.Conclusões. Percebe-se que a educação brindada pela universidade está diretamente relacionada ao desempenho do trabalho, estando satisfeitos com as competências de comunicação, investigação e processamento de informações

Propuesta de un sistema espacialmente explícito para evaluar el peligro de incendios

Los incendios forestales suponen un factor muy destacado en la transformación ambiental de buena parte de los ecosistemas terrestres. Tienen impactos globales, afectando a la superficie forestal y a las emisiones de gases de efecto invernadero, y efectos locales, relacionados con la degradación de suelos, erosión, modificación de la dinámica de la vegetación y pérdida de recursos y de vidas humanas. La prevención de incendios resulta cada vez más crítica, para paliar los efectos negativos de los mismos. Se presentan en este trabajo las variables de entrada y el esquema de integración para estimar el peligro de ocurrencia de incendios que se desarrolló en el marco del proyecto Firemap. Se generó información de diversas fuentes, que hacen referencia a variables socio-económicas, así como al estado de los combustibles y las características del territorio, utilizando sistemas de información geográfica (SIG) e imágenes de satélite. Todas las variables se cartografiaron a una resolución espacial de un 1 km2 y se integraron en un servidor web, que estuvo operativo para su evaluación durante el verano de 2007. Se presenta la comparación entre la variación temporal de los índices generados y la ocurrencia observada en la Comunidad de Madrid, una de las regiones del estudio.Forest fires are a major factor of environmental transformation in several ecosystems. Fires have global impacts, affecting forested areas and having an important impact in greenhouse gas emissions. Additionally, fires have local impacts, associated to soil degradation, soil erosion, vegetation dynamics, and lost of lives and properties. Fire prevention is critical to reduce the negative impacts of fire. This paper presents the input variables and the integration scheme developed within the Firemap project (funded under the Spanish Ministry of Science and Technology) to map wildland fire occurrence probability. The project first generated fire risk variables related to several factors of fire ignition or propagation. They were generated from a wide variety of sources using geographic information systems and remote sensing technologies. All variables were mapped at 1 sq km spatial resolution, and were integrated into single indices. The risk system included the development of a dedicated web-mapping server to facilitate the access to the end-users. This service was tested in the summer of 2007 for semi-operational use. The paper presents the first validation results of the danger index, by comparing temporal trends of the different danger components to the fire occurrence in the Madrid region, one of the test sites

Anexo II. Fichas de los ítems

El presente documento contiene uno de los anexos a la publicación impresa titulada "Recuperando la ciudad. Estrategia para el diseño y la evaluación de planes y programas de regeneración urbana integrada" —ISBN: 978-84-9728-5585-1— libro que es uno de los resultados obtenidos del proyecto financiado por el Plan Nacional I+D+i para el periodo 2013-2015, titulado "Estrategia para el diseño y evaluación de planes y programas de regeneración urbana integrada. La intervención en las periferias españolas a través de las áreas de rehabilitación integral y el programa URBAN" (BIA2012-31905). Dicho proyecto se ha realizado en el Departamento de Urbanística y Ordenación del Territorio de la Escuela Técnica Superior de Arquitectura de Madrid, mediante la financiación del Ministerio de Economía y Competitividad, a través del Plan Nacional de I+D+i 2008-2011 (Subprograma de Proyectos de Investigación Fundamental no Orientada). En este segundo anexo se realiza una caracterización y descripción de cada uno de los ítems que forman parte cada una de las categorías pertenecientes a las cuatro áreas principales, facilitando así el uso de la herramienta en los casos en los que se requiera de un mayor nivel de detalle

Recuperando la ciudad. Estrategia para el diseño y la evaluación de planes programas de regeneración urbana integrada

El presente documento es uno de los resultados obtenidos del proyecto financiado por el Plan Nacional I+D+i para el periodo 2013-2015, titulado Estrategia para el diseño y evaluación de planes y programas de regeneración urbana integrada. La intervención en las periferias españolas a través de las áreas de rehabilitación integral y el programa URBAN (BIA2012-31905), realizado en el Departamento de Urbanística y Ordenación del Territorio de la Escuela Técnica Superior de Arquitectura de Madrid. El objetivo del proyecto fue desarrollar una “herramienta de evaluación” de los planes y programas de Rehabilitación Urbana Integrada y se materializa en un árbol de conceptos que deberían ser analizados tanto antes como después del plan o proyecto. Su intención es facilitar la comunicación entre los distintos actores de la rehabilitación: vecinos, técnicos y administración, que al disponer de un modelo de análisis común (que deberá ser ratificado de forma consensuada desde las fases previas de la operación) puedan llegar a consensos sobre cuáles son los elementos fundamentales y los secundarios de la operación, lo que permitiría determinar (en un marco de recursos limitados), el porqué y las consecuencias de la elección de alguno de los elementos del modelo en detrimento de otros. El modelo se construye en una doble dimensión. La espacial en la que se reflexiona sobre los distintos niveles de integración, desde el área urbana en la que se enclava, al espacio público del barrio sobre el que se pretende actuar, para llegar a la edificación y la vivienda, incluyendo a los propios habitantes. Pero esta estructura tiene la capacidad de garantizar la segunda dimensión, la integralidad de las operaciones, diferenciándose ésta en tres niveles: áreas, categorías e ítems. Se definen cuatro áreas, Marco Urbano y Territorial, Diseño urbano y Medio ambiente local, Edificación y Socio-Económica, que establecen el marco básico de la calidad urbana. Las categorías, en las que se divide cada área, representan un marco conceptual consistente que establece los temas que tienen que ser considerados y analizados en el proceso si queremos garantizar la integralidad de la actuación. Las categorías se dividen en ítems que son los elementos que en cada caso se utilizarán para garantizar la calidad final del plan o programa. Se han considerado los ítems como contextuales porque se conformarán en detalle y con matices diferenciados en función de cada proyecto o acción concreta, por lo que podrán coincidir con los ítems elaborados durante el desarrollo de esta herramienta o ser diferentes por necesidades específicas. El presente documento se completa con dos anexos: Anexo I: Informe de la encuesta sobre experiencias de rehabilitación urbana en las ciudades españolas. Resultados. URL: http://oa.upm.es/43244/ Anexo II: Fichas de los ítems. URL: http://oa.upm.es/43247

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

Management of acute diverticulitis with pericolic free gas (ADIFAS). an international multicenter observational study

Background: There are no specific recommendations regarding the optimal management of this group of patients. The World Society of Emergency Surgery suggested a nonoperative strategy with antibiotic therapy, but this was a weak recommendation. This study aims to identify the optimal management of patients with acute diverticulitis (AD) presenting with pericolic free air with or without pericolic fluid. Methods: A multicenter, prospective, international study of patients diagnosed with AD and pericolic-free air with or without pericolic free fluid at a computed tomography (CT) scan between May 2020 and June 2021 was included. Patients were excluded if they had intra-abdominal distant free air, an abscess, generalized peritonitis, or less than a 1-year follow-up. The primary outcome was the rate of failure of nonoperative management within the index admission. Secondary outcomes included the rate of failure of nonoperative management within the first year and risk factors for failure. Results: A total of 810 patients were recruited across 69 European and South American centers; 744 patients (92%) were treated nonoperatively, and 66 (8%) underwent immediate surgery. Baseline characteristics were similar between groups. Hinchey II-IV on diagnostic imaging was the only independent risk factor for surgical intervention during index admission (odds ratios: 12.5, 95% CI: 2.4-64, P =0.003). Among patients treated nonoperatively, at index admission, 697 (94%) patients were discharged without any complications, 35 (4.7%) required emergency surgery, and 12 (1.6%) percutaneous drainage. Free pericolic fluid on CT scan was associated with a higher risk of failure of nonoperative management (odds ratios: 4.9, 95% CI: 1.2-19.9, P =0.023), with 88% of success compared to 96% without free fluid ( P &lt;0.001). The rate of treatment failure with nonoperative management during the first year of follow-up was 16.5%. Conclusion: Patients with AD presenting with pericolic free gas can be successfully managed nonoperatively in the vast majority of cases. Patients with both free pericolic gas and free pericolic fluid on a CT scan are at a higher risk of failing nonoperative management and require closer observation

La influencia de la tecnología en los procesos de producción musical: análisis de la grabación sonora y sus aplicaciones en el ámbito docente

Memoria de las actividades del proyecto de Innova-Docencia nº256, coordinado por el Dr. Marco Antonio Juan de Dios Cuartas, profesor del Departamento de Musicología de la Universidad Complutense de Madrid