Análisis de las proteínas nucleolares NopA100 y NopA64 en relación con la proliferación celular en Allium cepa L.

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid. Facultad de Medicina, Departamento de Bioquímica. Fecha de lectura: 21 de Abril 200

Involvement of raft aggregates enriched in Fas/CD95 death-inducing signaling complex in the antileukemic action of edelfosine in Jurkat cells

[Background]: Recent evidence suggests that co-clustering of Fas/CD95 death receptor and lipid rafts plays a major role in death receptor-mediated apoptosis. [Methodology/Principal Findings]: By a combination of genetic, biochemical, and ultrastructural approaches, we provide here compelling evidence for the involvement of lipid raft aggregates containing recruited Fas/CD95 death receptor, Fas-associated death domain-containing protein (FADD), and procaspase-8 in the induction of apoptosis in human T-cell leukemia Jurkat cells by the antitumor drug edelfosine, the prototype compound of a promising family of synthetic antitumor lipids named as synthetic alkyl-lysophospholipid analogues. Co-immunoprecipitation assays revealed that edelfosine induced the generation of the so-called death-inducing signaling complex (DISC), made up of Fas/CD95, FADD, and procaspase-8, in lipid rafts. Electron microscopy analyses allowed to visualize the formation of raft clusters and their co-localization with DISC components Fas/CD95, FADD, and procaspase-8 following edelfosine treatment of Jurkat cells. Silencing of Fas/CD95 by RNA interference, transfection with a FADD dominant-negative mutant that blocks Fas/CD95 signaling, and specific inhibition of caspase-8 prevented the apoptotic response triggered by edelfosine, hence demonstrating the functional role of DISC in drug-induced apoptosis. By using radioactive labeled edelfosine and a fluorescent analogue, we found that edelfosine accumulated in lipid rafts, forming edelfosine-rich membrane raft clusters in Jurkat leukemic T-cells. Disruption of these membrane raft domains abrogated drug uptake and drug-induced DISC assembly and apoptosis. Thus, edelfosine uptake into lipid rafts was critical for the onset of both co-aggregation of DISC in membrane rafts and subsequent apoptotic cell death. [Conclusions/Significance]: This work shows the involvement of DISC clusters in lipid raft aggregates as a supramolecular and physical entity responsible for the induction of apoptosis in leukemic cells by the antitumor drug edelfosine. Our data set a novel framework and paradigm in leukemia therapy, as well as in death receptor-mediated apoptosis.This work was supported by grants from Fondo de Investigacion Sanitaria and European Commission (FIS-FEDER 06/0813), Ministerio de Ciencia e Innovacion of Spain (SAF2008-02251 and RD06/0020/1037 - Red Tematica de Investigacion Cooperativa en Cancer, Instituto de Salud Carlos III), Junta de Castilla y Leon (CSI01A08, SAN673/SA32/08, and GR15 - Experimental Therapeutics and Translational Oncology Program), Fundacion “la Caixa” (BM05-30-0), and Fundacion de Investigacion Medica Mutua Madrilena (FMM). CG is supported by the Ramon y Cajal Program from the Ministerio de Ciencia e Innovacion of Spain.Peer Reviewe

Prevalence and Antimicrobial Susceptibility of Potentially Pathogenic Bacteria and Fungi Isolated from Public Parks of Bogotá, Colombia

Regular interactions of humans with their environment, animals and different microorganisms as part of everyday routine can cause several infections or diseases that can become a public health problem if these are not properly controlled. It is necessary to identify potentially pathogenic bacteria (with its antimicrobial resistance) and fungi, and their prevalence in public parks in Bogotá city. Four parks were evaluated, where samples were taken from dog’s feces and hair, recreational items and environmental material, to perform the microorganism’s isolations. The higher prevalence percentages in each sort of sample were for Escherichia coli, Staphylococcus epidermidis, Staphylococcus aureus, Rahnella aquatilis, Penicillium spp, Cladosporium spp. and Mucor spp. and there were pathogenic species like Salmonella enteritidis and Klebsiella pneumoniae. We report a strain of S. aureus with intermediate resistance to vancomycin (VISA) found in recreational items. The higher rates of antibiotics resistance like ampicillin and trimethoprim were found in the isolations from dog’s feces. Some species or genera reported are considerated as opportunistic pathogens; however, these and the pathogenic species reported represent a threat to human health and this is why it is essential to have a personal hygiene plan after stay at a park.Regular interactions of humans with their environment, animals and different microorganisms as part of everyday routine can cause several infections or diseases that can become a public health problem if these are not properly controlled. It is necessary to identify potentially pathogenic bacteria (with its antimicrobial resistance) and fungi, and their prevalence in public parks in Bogotá city. Four parks were evaluated, where samples were taken from dog’s feces and hair, recreational items and environmental material, to perform the microorganism’s isolations. The higher prevalence percentages in each sort of sample were for Escherichia coli, Staphylococcus epidermidis, Staphylococcus aureus, Rahnella aquatilis, Penicillium spp, Cladosporium spp. and Mucor spp. and there were pathogenic species like Salmonella enteritidis and Klebsiella pneumoniae. We report a strain of S. aureus with intermediate resistance to vancomycin (VISA) found in recreational items. The higher rates of antibiotics resistance like ampicillin and trimethoprim were found in the isolations from dog’s feces. Some species or genera reported are considerated as opportunistic pathogens; however, these and the pathogenic species reported represent a threat to human health and this is why it is essential to have a personal hygiene plan after stay at a park

Season-long seed dispersal patterns of the invasive weed Erigeron bonariensis in south-western Spain

Within-field demography of weeds exhibiting wind-mediated long distance seed movement can be largely governed by extra-field seed-source populations. Thus, for these species, a clear understanding of the temporal and spatial patterns of seed dispersal can benefit the development of effective management options. The spatial distribution of the seeds released from the onset of fruiting, in early summer, to the onset of the rainy season, in early autumn, was studied during 2 years at a Mediterranean-climate locality in Erigeron bonariensis L. (Hairy fleabane), a wind-dispersed invasive weed inhabiting ruderal environments and crop fields. Each year, a small source population was established in an open field in SW Spain and seed traps were arranged in the eight cardinal directions at distances up to 100 m (year 1), or in the NE and NW directions at distances up to 65 m (year 2). Counts of trapped seeds were carried out at 4–6 day intervals and the number of seeds released by the source population was estimated each year in most census dates. Four empirical dispersal models based on either thin-tailed or fat-tailed density kernels were tested using year 1 data for their ability to represent the spatial distribution of seeds. To test for anisotropic dispersal, model parameters were allowed to vary according to the wind pattern in each cardinal direction. Based on information criteria, a model including a fat-tailed, Log-hyperbolic secant kernel showing parameter response to the wind pattern, highlighting striking anisotropic dispersal, was selected and evaluated using year 2 data. Distance percentiles 50 and 80 attained by the seed crop released in year 1 season were modeled at 530 m and 10,498 m, respectively. The opposite quadrants encompassing the dominant downwind (N-NE) and upwind directions (S-SW) received 52.5% and 10.8% of seeds. The year 1 population, consisting of 85 plants, generated a modeled seed rain of at least 10 seeds m 2 up to 200 m downwind. Implications of results for management of this herbicide resistance-prone species are discussed.We are grateful to Carlos Carretero, Jos´e P´erez, C´ andido Ortega and Ana Lebovich for assistance with fieldwork. DC received a grant from the Spanish Society of Weed Science (SEMh). This work was supported in part by research project C03-017 (PAI-Junta de Andalucía Regional Government, Andalusia, Spain). Funding for open access provided by Universidad de Huelva / CBU

Clinical Relevance and Molecular Pathogenesis of the Emerging Serotypes 22F and 33F of Streptococcus pneumoniae in Spain

Streptococcus pneumoniae is the main bacterial cause of respiratory infections in children and the elderly worldwide. Serotype replacement is a frequent phenomenon after the introduction of conjugated vaccines, with emerging serotypes 22F and 33F as frequent non-PCV13 serotypes in children and adults in North America and other countries. Characterization of mechanisms involved in evasion of the host immune response by these serotypes is of great importance in public health because they are included in the future conjugated vaccines PCV15 and PCV20. One of the main strategies of S. pneumoniae to persistently colonize and causes infection is biofilm formation. In this study, we have evaluated the influence of capsule polysaccharide in biofilm formation and immune evasion by using clinical isolates from different sources and isogenic strains with capsules from prevalent serotypes. Since the introduction of PCV13 in Spain in the year 2010, isolates of serotypes 22F and 33F are rising among risk populations. The predominant circulating genotypes are ST43322F and ST71733F , being CC433 in 22F and CC717 in 33F the main clonal complexes in Spain. The use of clinical isolates of different origin, demonstrated that pediatric isolates of serotypes 22F and 33F formed better biofilms than adult isolates and this was statistically significant. This phenotype was greater in clinical isolates from blood origin compared to those from cerebrospinal fluid, pleural fluid and otitis. Opsonophagocytosis assays showed that serotype 22F and 33F were recognized by the PSGL-1 receptor on leukocytes, although serotype 22F, was more resistant than serotype 33F to phagocytosis killing and more lethal in a mouse sepsis model. Overall, the emergence of additional PCV15 serotypes, especially 22F, could be associated to an enhanced ability to divert the host immune response that markedly increased in a biofilm state. Our findings demonstrate that pediatric isolates of 22F and 33F, that form better biofilm than isolates from adults, could have an advantage to colonize the nasopharynx of children and therefore, be important in carriage and subsequent dissemination to the elderly. The increased ability of serotype 22F to avoid the host immune response, might explain the emergence of this serotype in the last years.This work was partially supported by a grant from the MSD-USA (MISP Call, Reference: 57320), Ministerio de Economía, Industria y Competitividad (MINECO) (SAF2017-83388-R), and CIBER de Enfermedades Respiratorias (CIBERES) is an initiative of the Instituto de Salud Carlos III

A GRASP-Tabu Heuristic Approach to Territory Design for Pickup and Delivery Operations for Large-Scale Instances

Weaddressalogisticsdistrictingproblemfacedbyaparcelcompanywhoseoperationsconsistofpickingupanddeliveringpackages overaserviceregion.Thedistrictingprocessaimstofindapartitionoftheserviceregionintodeliveryandcollectionzonesthat may be served by a single vehicle that departs from a central depot. Criteria to be optimized are to balance workload content among the districts and to create districts of compact shape. A solution approach based on a hybrid procedure that combines elements of GRASP and Tabu Search (TS) is proposed to solve large-scale instances. Numerical experimentation is performed consideringdifferentinstancesizesandtypes.Resultsshowthattheproposedsolutionapproachisabletosolvelarge-scaleinstances inreasonablecomputationaltimeswithgoodqualityofthesolutionsobtained.Todeterminethequalityofthesolutions,resultsare comparedwithCPLEXsolutionsandwiththecurrentrealsolutiontohighlightthebenefitsoftheproposedapproach.Conclusions andrecommendationsforfurtherresearchareprovided

Knowledge management model in the productive chain of health services providing entities

En el presente artículo se formula y presenta un modelo de gestión del conocimiento adecuado con base en el modelo dinámico de creación del conocimiento de Nonaka y Takeuchi, para el mejoramiento de la cadena productiva de las entidades prestadoras de salud con aseguramiento de la calidad en los servicios de atención al usuario, teniendo en cuenta la importancia de los mismos, y la ineficiencia que se presenta al acceder a ellos, para ello se efectúa el diagnóstico y análisis de la calidad del servicio en salud, teniendo en cuenta que el resultado del mismo se encuentra relacionado con la eficiencia de los eslabones y actores de la cadena productiva, la cual brinda los parámetros para la creación del modelo de GC, la arquitectura y por ende su respectiva adaptación. Posterior a ello se mencionan dos herramientas multidimensionales de medición de la calidad del servicio, SERVQUAL para la medición y evaluación de la calidad del servicio al cliente brindado a los usuarios y SERVQHOS la evaluación de la percepción de la calidad del servicio, con los cuales las entidades prestadoras de salud podrán evaluar, y analizar la efectividad del modelo propuesto y los procesos a mejorar en la cadena productiva y así mismo fomentar la creación de una permanente sociedad del conocimiento en la organización.In this article, an adequate knowledge management model is formulated and presented based on the dynamic model of knowledge creation of Nonaka and Takeuchi, for the improvement of the productive chain of health providers with quality assurance in the user care services, taking into account their importance, and the inefficiency that occurs when accessing them, for this, the diagnosis and analysis of the quality of the health service is carried out, taking into account that the result of the same it is related to the efficiency of the links and actors of the production chain, which provides the parameters for the creation of the CG model, the architecture and therefore its respective adaptation. After this, two multidimensional tools for measuring the quality of the service are mentioned, SERVQUAL for the measurement and evaluation of the quality of the customer service provided to users and SERVQHOS the evaluation of the perception of the quality of the service, with which the Health provider entities will be able to evaluate and analyze the effectiveness of the proposed model and the processes to be improved in the production chain and also promote the creation of a permanent knowledge society in the organization

Evaluación de un programa de políticas activas de mercado de trabajo en un contexto de elevado desempleo

We evaluate the effectiveness of a programme aimed at a group of unemployed in the capital of the South of Spain, within the framework of Active Labour Market Policies (almps). We use high quality administrative data which justifies the application of propensity score matching methods. The estimated effects are positive with regard to employment, job security, and working hours in the short-term (6 months). However, this is not true in the long-run (36 months). No significant effects have been found on earnings, in neither the short nor long-term. Overall these results are quite robust with respect to the matching algorithm choice and the potential influence of unobserved heterogeneity. Although, the short duration of the programme seems appropriate, the disappointing long-term results highlight the difficulties of putting participants back into stable work in a context of high unemploymentEste trabajo evalúa el impacto de un programa dirigido a un grupo de desempleados en la capital del sur de España, en el marco de las políticas activas del mercado de trabajo (pamt). Se han utilizado para ello datos administrativos de alta calidad, lo que justifica la aplicación de métodos de propensity score matching (psm). El efecto estimado es positivo en materia de empleo, seguridad en el empleo y horas de trabajo, en el corto plazo (6 meses), pero no en el largo plazo (36 meses). No se ha encontrado un efecto significativo en los ingresos, ni en el corto ni en el largo plazo. En general, estos resultados son bastante consistentes con respecto al algoritmo matching elegido y la influencia potencial de la heterogeneidad inobservada. Aunque la corta duración del programa parece apropiada, los pobres resultados a largo plazo reflejan las dificultades de los participantes para conseguir un trabajo estable en un contexto de elevado desemple

Clonal dynamics in osteosarcoma defined by RGB marking

Osteosarcoma is a type of bone tumour characterized by considerable levels of phenotypic heterogeneity, aneuploidy, and a high mutational rate. The life expectancy of osteosarcoma patients has not changed during the last three decades and thus much remains to be learned about the disease biology. Here, we employ a RGB-based single-cell tracking system to study the clonal dynamics occurring in a de novo-induced murine osteosarcoma model. We show that osteosarcoma cells present initial polyclonal dynamics, followed by clonal dominance associated with adaptation to the microenvironment. Interestingly, the dominant clones are composed of subclones with a similar tumour generation potential when they are re-implanted in mice. Moreover, individual spontaneous metastases are clonal or oligoclonal, but they have a different cellular origin than the dominant clones present in primary tumours. In summary, we present evidence that osteosarcomagenesis can follow a neutral evolution model, in which different cancer clones coexist and propagate simultaneously.We thank ISCIII and CNIO flow cytometry and cell sorting units for their participation in our studies. We are thankful to the CCEH-Fred Hutchinson Cancer Research Center for LAM-PCR service. We acknowledge Raquel Pérez Tavarez, María Blázquez Mesa, Alicia Giménez Sánchez, Elena Calvo Cazalilla, and Monserrat Arroyo Correas for useful help on the pathology studies; and Teresa Cejalvo, Isabel Cubillo Moreno, and Miguel Angel Rodríguez-Milla for their contributions in experimental setup. We thank the visual artist Isabella Lacquaniti for her help with drawings and schematics. We are also thankful to the Fondo de Investigaciones Sanitarias (FIS: PI11/00377 and PI14CIII/00005 to J.G.-C., FIS: CP11/00206 to A.A., and RTICC: RD12/0036/0027 to J.G.-C.), the Madrid Regional Government (CellCAM; P2010/BMD-2420 to J.G.-C.), the Asociación Pablo Ugarte, and the Asociación Afanion for grants support.S

VALIDAÇÃO DE UMA ESCALA PARA A ANÁLISE DAS PERCEPÇÕES DOS MORADORES SOBRE O IMPACTO SOCIAL DE UM CENTRO ESPORTIVO

The purpose of this study is to validate a scale for the analysis of the social impact of a sports center. A sample of 406 residents of Moncada (Valencia) was collected with a sample error of ± 4.82, who answered a survey of 31 items that collected possible social impacts derived from the presence of the sports center in the town. The implementation of exploratory and confirmatory factor analysis reduced the scale to 28 indicators distributed in seven dimensions of impact: socio-cultural impact, socio-economic impact, impact on image and promotion of the town, impact on urban development and infrastructures, impact on social cohesion, equality and equity, impact on health and impact on habits and levels of physical activity. The results allowed to verify the validity and reliability of the scale proposed for the object of study.La finalidad de este estudio es validar una escala para el análisis de las percepciones de los residentes sobre el impacto social de un centro deportivo. Se recogió una muestra de 406 residentes del municipio de Moncada (Valencia), con un error de muestreo de ±4,82, que contestaron a una encuesta de 31 ítems que recogían posibles impactos sociales derivados de la presencia del centro deportivo en la localidad. La aplicación de análisis factorial exploratorio y confirmatorio redujo la escala a 28 indicadores distribuidos en siete dimensiones de impacto: impacto sociocultural, impacto socioeconómico, impacto en la imagen y la promoción del municipio, impacto en el desarrollo urbano y las infraestructuras, impacto en la cohesión social, igualdad y equidad, impacto sobre la salud e impacto sobre los hábitos y niveles de actividad física. Los resultados permitieron comprobar la validez y fiabilidad de la escala propuesta para el objeto de estudio. O objetivo deste estudo foi validar uma escala para a análise das percepções dos moradores sobre o impacto social de um centro esportivo. Se recolheu uma amostra de 406 moradores do município de Moncada (Valência) com erro de amostragem de ± 4,82. Os moradores responderam a um questionário de 31 itens que incluíram possíveis impactos sociais derivados da presença do complexo esportivo na localidade. A aplicação da análise fatorial exploratória e confirmatória reduziu a escala para 28 indicadores distribuídos em sete dimensões de impacto: impacto sociocultural, impacto socioeconômico, impacto na imagem e promoção do município, impacto no desenvolvimento e infraestruturas urbanas, impacto na coesão social, igualdade e equidade, impacto na saúde e impacto nos hábitos e níveis de atividade física. Os resultados permitiram verificar a validade e confiabilidade da escala proposta para o objeto de estudo.