Poblaciones de hongos toxicogénicos y fermentación ruminal in vitro en granos de sorgo ensilados : efectos de la variedad del grano, contenido de humedad y tiempo de almacenamiento

El objetivo de la presente tesis de doctorado fue evaluar el efecto de la variedad, el conte-nido de humedad y el tiempo de almacenamiento sobre las poblaciones fúngicas toxicogé-nicas y los parámetros de fermentación ruminal in vitro en granos de sorgo ensilados hú-medos. Se realizaron tres estudios experimentales, el primero en condiciones de campo en donde se obtuvieron muestras de cuatro establecimientos lecheros comerciales al momento de la cosecha de granos de sorgo de variedades altas y bajas en concentraciones de taninos, almacenados húmedos en silo-bolsas. Los granos de estos silos fueron evaluados al inicio (tiempo 0) y a los 30, 90 y 180 días de almacenamiento. En estas muestras se analizaron la composición química, los parámetros de fermentación in vitro y las poblaciones fúngicas totales y las toxicogénicas. Los resultados de este estudio sugieren que 180 días de alma-cenamiento fue beneficioso al reducir la concentración de taninos condensados, aumentan-do la velocidad de fermentación de los granos y minimizando la contaminación fúngica, tanto en las variedades altas como bajas en taninos. Ya a los 90 días se vieron las condicio-nes óptimas para minimizar la contaminación con hongos toxicogénicos. Los otros dos estudios, fueron realizados en condiciones de laboratorio, a partir de las muestras de granos de sorgo colectados al momento de la cosecha de los establecimientos del primer estudio más un quinto establecimiento. Estas muestras fueron almacenadas en silos experimenta-les, recibieron tres tratamientos de humedad diferentes y fueron evaluadas a los 30, 90 y 180 días de almacenamiento. En el segundo estudio se analizaron hongos toxicogénicos y micotoxinas y en el tercero la fermentación ruminal in vitro. Los resultados del segundo estudio evidenciaron una baja contaminación por hongos toxicogénicos y micotoxinas. En tanto que la colonización de Penicillium spp. se vio favorecida en los granos almacenados en relación con los granos frescos antes del ensilaje. Las poblaciones de Fusarium spp. disminuyeron después de los 30 días de almacenamiento lo mismo que las de Aspergillus spp. en granos con alto contenido de taninos. En el tercer estudio, los granos de sorgo de variedad baja en taninos produjeron un mayor volumen de gas. Los granos de sorgo de variedad alta en taninos fermentaron mejor con un contenido de humedad medio, en cam-bio, los granos bajos en taninos fermentaron mejor con un contenido de humedad alto. Tanto los granos de sorgo altos como los bajos en taninos mostraron una mejor fermenta-ción ruminal con un mayor tiempo de almacenamiento. Los resultados de estos estudios sugieren que granos de sorgo de variedades altas o bajas en taninos, ensilados con un con-tenido de humedad entre 26 y 32% y almacenados por un tiempo mínimo de 90 días, ópti-mo 180 días, serían las condiciones ideales que favorecen la fermentiscibilidad de los gra-nos y minimizan la contaminación fúngica y de micotoxinas. Es importante realizar un monitoreo periódico del ensilaje de sorgo en diferentes momentos de acuerdo con la evolu-ción de la población de hongos y la producción de micotoxinas

Efecto citotóxico de extractos de Senecio madagascariensis (Asteraceae) en lineas celulares derivadas de cáncer

Species of the genus Senecio have been traditionally employed with medical purposes and it has been demonstrated that some of them have anti-proliferative activity on cancer-derived cell lines. In South America S. madagascariensis is one of the most important representatives of the genus. Almost all species contain pyrrolizidine alkaloids (PA), substances that can lead to hepatic damage. Thus, the aim of this work was to determine the cytotoxic activity of a low PAs extract of S. madagascariensis on a cancer-derived cell line. A chloroform extract from plant material was produced and it was demonstrated to have a PAs concentration lower than 200 µg/g (w/w). Two stock solutions were made with DMSO and acetone as solvents and were diluted in culture medium (DMEM). Cells from urinary bladder transitional cell carcinoma T24 were cultured in 96 well plates (10⁴ cells/well) and incubated for 24 hours with dilutions of the extracts at the following concen-trations, 0.5 mg/mL, 0.25 mg/mL, 0.125 mg/mL and 0.06 mg/mL. Cell viability was evaluated by the MTT col-orimetric technique. Both solutions of S. madagascariensis extracts (acetone and DMSO) showed cytotoxic activity. In both cases, the degree of cytotoxicity was dependent on the concentration of the solution.Las especies del género Senecio han sido empleadas tradicionalmente con fines medicinales y se ha demos-trado que algunas de ellas poseen actividad antiproliferativa en células cancerosas. En América del Sur, S. madagascariensis es uno de los representantes más importantes del género. La mayoría de las especies contienen alcaloides pirrolizidínicos (AP), sustancias que pueden provocar daño hepático. Por lo tanto, el obje-tivo de este trabajo fue determinar la actividad citotóxica de un extracto bajo en AP de S. madagascariensis en una línea celular cancerosa. Se desarrolló un extracto clorofórmico a partir del material vegetal con una concentración de AP menor a 200 µg/g (w/w). Dos soluciones stock fueron realizadas utilizando DMSO y acetona como solventes y fueron diluidas en medio de cultivo (DMEM). Fueron cultivadas células de carcino-ma de células transicionales de vejiga urinaria T24 en placas de 96 pocillos (10⁴ células por pocillo) e incuba-das por 24 horas con diluciones de los extractos a las siguientes concentraciones: 0,5 mg/mL, 0,25 mg/mL, 0,125 mg/mL y 0,06 mg/mL. La viabilidad celular fue evaluada mediante la técnica colorimétrica de MTT. Am-bas soluciones de S. madagascariensis demostraron actividad citotóxica. En ambos casos el grado de citoto-xicidad fue dependiente de la concentración de la solución.As especies do gênero Senecio têm sido tradicionalmente utilizadas para fins medicinais e algumas de las mostraram ter atividade antiproliferativa em células cancerosas. Em América do Sul, S. madagascariensis é um dos representantes mais importantes do gênero. A maioria das espécies contém alcalóides pirrolizidínicos (PAs), substâncias que podem causar dano ao fígado. Portanto, o objetivo deste trabalho foi determinar a atividade citotóxica de um extrato de S. madagascariensis com baixo teor de PA em uma linhagem de células cancerosas. Um extrato clorofórmico foi realizado a partir do material vegetal com uma concentração de PA inferior a 200 µg/g (w/w). Dois soluções estoque foram feitas usando DMSO e acetona como solventes e foram diluídas em meio de cultura (DMEM). Células de carcinoma de células transicionais da bexiga urinária T24 foram cultivadas em placas de 96 poços (10⁴ células por poço) e incubadas por 24 horas com diluições dos extratos nas seguintes concentrações 0,5 mg / mL, 0,25 mg / mL, 0,125 mg / mL e 0,06 mg / mL. A viabi-lidade celular foi avaliada pela técnica colorimétrica MTT. Ambas soluções de S. madagascariensis demons-traram atividade citotóxica. Em ambos casos, o grau de citotoxicidade foi dependente da concentração da solução

Standards and specifications to manage accessibility issues in e-learning

Despite the acknowledged need of providing a personalized and adaptive learning process for all, current learning management systems do not properly cover personalization and accessibility issues and they are still struggling to support the reusability requirements coming from the pervasive usage of standards. There is a lack of frameworks for providing layered-based infrastructure covering the interoperability required to manage the whole range of standards, applications and services needed to meet accessibility and adaptations needs of lifelong learning services

Vitamin D nutritional status and vitamin D regulated antimicrobial peptides in serum and pleural fluid of patients with infectious and noninfectious pleural effusions

Background: Vitamin D and vitamin D dependent antimicrobial peptides such as Cathelicidin (LL-37) and ?-defensin 2 have an important role in innate and adaptative immunity, but their role in pleural effusions has not been studied before. Methods: Serum and pleural fluid samples from 152 patients with pleural effusion were collected, corresponding to 45 transudates and 107 exudates, 51 infectious effusions (14 complicated and 37 non-complicated), 44 congestive heart failure effusions and 38 malignant effusions. The levels of 25 OH-vitamin D, 1,25-(OH)2-vitamin D, Vitamin D Binding Protein (VDBP), LL-37 and ?-defensin 2, both in serum and pleural fluid were evaluated in this prospective study. Differences between groups were analysed using unpaired t tests or Mann?Whitney tests. Correlations between data sets were examined using Pearson correlation coefficient or Spearman rank correlation coefficient. Diagnostic accuracy was estimated using ROC curve analysis. Results: Low serum 25 OH vitamin D levels were found in all groups. Infectious effusions (IE) had higher serum and pleural fluid LL-37 levels compared to congestive heart failure or malignant effusions. Among IE, complicated had higher serum and pleural fluid LL-37 levels, and lower serum ?-defensin-2 levels. Positive correlations were found between serum 25 OH-vitamin D levels and serum or pleural 1,25-(OH)2-vitamin D levels, and between 1,25-(OH) 2-vitamin D and LL-37 serum. Diagnostic accuracy of the different molecules was moderate at best. Conclusions: Hypovitaminosis D is highly prevalent in pleural effusions. LL-37 is produced intrapleurally in IE. This production is higher in complicated IE. No evidence of pleural production of ?-defensin 2 was found in any of the groups. Diagnostic accuracy of the different molecules is at the best moderate for discriminating different types of effusions

Marco activo de recursos de innovación docente: Madrid

Una guía de espacios e instituciones para actividades educativas complementarias en enseñanza secundaria y Formación Profesional

SMAD3 rs17228212 Gene Polymorphism Is Associated with Reduced Risk to Cerebrovascular Accidents and Subclinical Atherosclerosis in Anti-CCP Negative Spanish Rheumatoid Arthritis Patients

Rheumatoid arthritis (RA) is a complex polygenic inflammatory disease associated with accelerated atherosclerosis and increased risk of cardiovascular (CV) disease. Previous genome-wide association studies have described SMAD3 rs17228212 polymorphism as an important signal associated with CV events. The aim of the present study was to evaluate for the first time the relationship between this gene polymorphism and the susceptibility to CV manifestations and its potential association with the presence of subclinical atherosclerosis assessed by the evaluation of carotid intima-media thickness (cIMT) in patients with RA

Targeting of PI3K/AKT/mTOR pathway to inhibit T cell activation and prevent graft-versus-host disease development

Producción CientíficaBackground: Graft-versus-host disease (GvHD) remains the major obstacle to successful allogeneic hematopoietic stem cell transplantation, despite of the immunosuppressive regimens administered to control T cell alloreactivity. PI3K/AKT/mTOR pathway is crucial in T cell activation and function and, therefore, represents an attractive therapeutic target to prevent GvHD development. Recently, numerous PI3K inhibitors have been developed for cancer therapy. However, few studies have explored their immunosuppressive effect. Methods: The effects of a selective PI3K inhibitor (BKM120) and a dual PI3K/mTOR inhibitor (BEZ235) on human T cell proliferation, expression of activation-related molecules, and phosphorylation of PI3K/AKT/mTOR pathway proteins were analyzed. Besides, the ability of BEZ235 to prevent GvHD development in mice was evaluated. Results: Simultaneous inhibition of PI3K and mTOR was efficient at lower concentrations than PI3K specific targeting. Importantly, BEZ235 prevented naïve T cell activation and induced tolerance of alloreactive T cells, while maintaining an adequate response against cytomegalovirus, more efficiently than BKM120. Finally, BEZ235 treatment significantly improved the survival and decreased the GvHD development in mice. Conclusions: These results support the use of PI3K inhibitors to control T cell responses and show the potential utility of the dual PI3K/mTOR inhibitor BEZ235 in GvHD prophylaxis.Asociación Española Contra el Cáncer (Proyecto AIOA110296BLAN).Gerencia Regional de Salud de Castilla y León (Proyecto GRS 726/A13

A Functional Role of RB-Dependent Pathway in the Control of Quiescence in Adult Epidermal Stem Cells Revealed by Genomic Profiling

Continuous cell renewal in mouse epidermis is at the expense of a pool of pluripotent cells that lie in a well defined niche in the hair follicle known as the bulge. To identify mechanisms controlling hair follicle stem cell homeostasis, we developed a strategy to isolate adult bulge stem cells in mice and to define their transcriptional profile. We observed that a large number of transcripts are underexpressed in hair follicle stem cells when compared to non-stem cells. Importantly, the majority of these downregulated genes are involved in cell cycle. Using bioinformatics tools, we identified the E2F transcription factor family as a potential element involved in the regulation of these transcripts. To determine their functional role, we used engineered mice lacking Rb gene in epidermis, which showed increased expression of most E2F family members and increased E2F transcriptional activity. Experiments designed to analyze epidermal stem cell functionality (i.e.: hair regrowth and wound healing) imply a role of the Rb-E2F axis in the control of stem cell quiescence in epidermis

Non-motor symptom burden in patients with Parkinson's disease with impulse control disorders and compulsive behaviours : results from the COPPADIS cohort

The study was aimed at analysing the frequency of impulse control disorders (ICDs) and compulsive behaviours (CBs) in patients with Parkinson's disease (PD) and in control subjects (CS) as well as the relationship between ICDs/CBs and motor, nonmotor features and dopaminergic treatment in PD patients. Data came from COPPADIS-2015, an observational, descriptive, nationwide (Spain) study. We used the validated Questionnaire for Impulsive-Compulsive Disorders in Parkinson's Disease-Rating Scale (QUIP-RS) for ICD/CB screening. The association between demographic data and ICDs/CBs was analyzed in both groups. In PD, this relationship was evaluated using clinical features and treatment-related data. As result, 613 PD patients (mean age 62.47 ± 9.09 years, 59.87% men) and 179 CS (mean age 60.84 ± 8.33 years, 47.48% men) were included. ICDs and CBs were more frequent in PD (ICDs 12.7% vs. 1.6%, p < 0.001; CBs 7.18% vs. 1.67%, p = 0.01). PD patients had more frequent previous ICDs history, premorbid impulsive personality and antidepressant treatment (p < 0.05) compared with CS. In PD, patients with ICDs/CBs presented younger age at disease onset, more frequent history of previous ICDs and premorbid personality (p < 0.05), as well as higher comorbidity with nonmotor symptoms, including depression and poor quality of life. Treatment with dopamine agonists increased the risk of ICDs/CBs, being dose dependent (p < 0.05). As conclusions, ICDs and CBs were more frequent in patients with PD than in CS. More nonmotor symptoms were present in patients with PD who had ICDs/CBs compared with those without. Dopamine agonists have a prominent effect on ICDs/CBs, which could be influenced by dose