In vitro study of CD133 human stem cells labeled with superparamagnetic iron oxide nanoparticles

Superparamagnetic iron oxide nanoparticles (SPIONs) are applied in stem cell labeling because of their high magnetic susceptibility as compared with ordinary paramagnetic species, their low toxicity, and their ease of magnetic manipulation. The present work is the study of CD133(+) stem cell labeling by SPIONs coupled to a specific antibody (AC133), resulting in the antigenic labeling of the CD133+ stem cell, and a method was developed for the quantification of the SPION content per cell, necessary for molecular imaging optimization. Flow cytometry analysis established the efficiency of the selection process and helped determine that the CD133 cells selected by chromatographic affinity express the transmembrane glycoprotein CD133. The presence of antibodies coupled to the SPION, expressed in the cell membrane, was observed by transmission electron microscopy. Quantification of the SPION concentration in the marked cells using the ferromagnetic resonance technique resulted in a value of 1.70 x 10 (13) mol iron (9.5 pg) or 7.0 x 10 (6) nanoparticles per cell ( the measurement was carried out in a volume of 2 mu L containing about 6.16 x 10 5 pg iron, equivalent to 4.5 x 10 (11) SPIONs). (c) 2008 Elsevier Inc. All rights reserved.

Expression of Extracellular Matrix Proteins in Human Dental Pulp Stem Cells Depends on the Donor Tooth Conditions

Introduction: Stem cells are characterized by the ability to renew themselves through mitotic cell division and differentiating into a diverse range of specialized cell types. An important source of adult stem cells is the dental pulp. In dentistry, regenerative strategies are of importance because of hard dental tissue damage especially as result of caries lesions, trauma, or iatrogenic procedures. The regeneration of dental tissues relies on the ability of stem cells to produce extracellular (ECM) proteins encountered in the dental pulp tissue. Thus, the aim of this study was to analyze the expression and distribution of proteins encountered in dental pulp ECM (type I collagen, fibronectin, and tenascin) in stem cells. Methods: Human immature dental pulp stem cells (hIDPSCs) from deciduous (DL-1 and DL-4 cell lines) and permanent (DL-2) teeth were used. The distribution of ECM proteins was observed using the immunofluorescence technique. The gene expression profile was evaluated using reverse transcription polymerase chain reaction (RT-PCR) analysis. Results: Positive reactions for all ECM proteins were observed independently of the hIDPSCs analyzed. Type I collagen appeared less evident in DL-2 than in other hIDPSCs. Fibronectin and tenascin were less clear in DL-4. The RT-PCR reactions showed that type I collagen was lesser expressed in the DL-2 cells, whereas fibronectin and tenascin were similarly expressed in all hIDPSCs. Conclusions: The distribution and expression of ECM proteins differ among the hIDPSCs. These differences seemed to be related to the donor tooth conditions (deciduous or permanent, retained or erupted, and degree of root reabsorption). (J Endod 2010;36:826-831)

The relationship between leaf water potential and stem diameter in sorghum

Leaf water potential (psi (l)) represents a good indicator of the water status of plants, and continuous monitoring of it can be useful in research and field applications such as scheduling irrigation. Changes in stem diameter (Sd) were used for monitoring psi (l) of pot-grown sorghum [Sorghum bicolor (L.) Moench] plants in a glasshouse. This method requires occasional calibration of S-d values against psi (l). Predicted values of psi (l), based on a single calibration show a good correlation with measured psi (l), values over a period of 13 d before and after the calibration. The correlation can further be improved with shorter time intervals.

Trends in physical activity participation and the impact of integrated campaigns among Australian adults, 1997-99

Objectives: To determine whether awareness of the moderate physical activity message and prevalence of participation changed among Australian adults between 1997 and 1999, and if changes differed across Australia. Methods: Data were compared on awareness of the moderate physical activity message and on physical activity participation from identical national physical activity surveys in 1997 and 1999. Results: In 1999, following integrated public health efforts, recognition of the Active Australia campaign was substantially higher in NSW/ACT (61.7%) than elsewhere (29.3%). Knowledge about benefits of moderate activity increased between 1997 and 1999, more so in States with public health campaigns. National participation in 'sufficient physical activity' declined between 1997 and 1999, from 63% to 57%, but the decline was smaller in NSW/ACT (4.4%) than in the other States (6.0%). Conclusions and implications: Declining trends in physical activity in Australia require increased public health investments, including strategic planning and public education, such as occurred in NSW (1997/98).

Public Participation geographic information systems in developing countries: lessons from Uganda on the impacts of interactive screens PPGIS and community perceptions of space

Dissertation submitted in partial fulfillment of the requirements for the Degree of Master of Science in Geospatial Technologies.

Radio mobility in the digital era: interactivity, participation and content share possibilities in Iberian broadcasters

In the context of the scientific research into radio, recent years have encouraged many theories about the meaning of a post-radio (Oliveira & Portela, 2011), thus enlisting several parameters regarding the inclusion of contemporary radio in the digital and online environments. This digital migration has led to the development of mobile applications for radio, broadening the communicative potential of audiences (Aguado, Feijoo & Martínez, 2013), as well as promoting convergence of interactive content among listeners-users. Aware of this opportunity, the main broadcasters in Spain and Portugal have broadened their radiophonic scope to the mobile platform, especially geared towards smartphones through the development of mobile applications, commonly known as apps (Cerezo, 2010). As a symbol of a culture in permanent changing, smartphones not only provide greater easiness in terms of access and interaction, but also afford larger opportunities for disseminating content among audiences, a phenomenon that some studies have labelled as user distributed content (Villi, 2012). This article presents an exploratory analysis of the current policies of the main Spanish and Portuguese radio broadcasters regarding mobile applications, evaluating the different levels of interaction and participation in these platforms. This observation led to the conclusion, among other findings, that the mobile platform represents a supplementary channel for traditional FM radio, rather than a new medium with its own language and expression.

Listening to the public: enacting power : citizen access, Standing and influence in public participation discourses

Public participation in environmental governance is typically associated with citizen access to power despite many closures and limitations having been identified in participatory processes. This article proposes an analytical framework to analyse discursive practices involved in public consultation processes. Critical Discourse Analysis is used to examine and appraise citizens’ access, standing and influence. We apply that framework to a ‘notice and comment’ process on a hydroelectric power plan in Portugal and show that it was discursively managed to justify the decision of constructing 10 large dams and to reject critical or alternative views. Citizens’ access, standing and influence were constrained through diverse discursive practices which (re)produced very unequal power relationsbetween policy proponents and participating individuals. More generally, the article illustrates the potential of Critical Discourse Analysis to assess voice(s) in policy processes. Focusing on argumentative, interactional and rhetorical levels, and how they are interwoven in public consultation discourses, the proposed framework is conceivably applicable in other studies.

Derechos de participación e información en programas dirigidos a adolescentes en escuelas secundarias de la Ciudad de Córdoba: cumplimiento progresivo y estrategias de promoción. Participation and information rights in programs for adolescents in secondary schools in Cordoba City: progressive implementation and promotion strategies.

Abordar la infancia desde el enfoque de derechos (ED) implica una lectura de los derechos de los cuales los niños, niñas y adolescentes (NNA) son titulares y de las acciones estatales que se implementan para viabilizar el ejercicio concreto de ellos. Esta perspectiva exige comprender los contenidos de las políticas púb,el grado de participación que se les asigna y los sentidos que ellas toman en términos de progresividad de los derechos. Los cambios de concepción de la infancia desde la Convención Internacional de los Derechos del Niño (CIDN) y los cambios legislativos interpelan al Estado y la sociedad a modificar la visión hegemónica. Nuestra hipótesis gira alrededor de las siguientes considerac.: se han producido algunas modificaciones en las políticas y acciones estatales, aún no se dan las transformaciones necesarias de las lógicas institucionales y de los operadores que estimulen la participación de NNA en el ámbito escolar. Esto se visualiza en la ausencia de análisis cuali de los contenidos de las acc. y la falta de ftes estadísticas específicas, que muestren la orientac.de los contenidos en los principios de la protecc.integral. Estos presupuestos plantean interrogantes que delimitan el problema de investigación: ¿cómo se construye y reconstruye el ejercicio de la ciudadanía a través de la participación de los NNA, en su paso por la escuela, y cuáles son las lógicas institucionales que habilitan u obstaculizan ese ejercicio? Y ¿cuáles son los recursos institucionales que permitan que los programas y acciones redunden en el ejercicio efectivo de los derechos?.Obj.gral: Describir, analizar e interpretar la progresividad en el ejercicio de los derechos de participación e informacióin que tienen los adolescentes en escuelas secundarias de la cd de Cba, durante 2005 / 2010, para promover espacios públicos de construcción de ciudadanía. Se analizarán tres escuelas de Córdoba, escuela pública provincial, púb. nacional y privada. Obj. Espec.: Analizar las prácticas participativas formales e informales en las escuelas seleccionadas.- Aplicar indicadores cuanti y cualitativos del “Sistema de Indicadores para los derechos de la Infancia” Describir y analizar los recursos materiales y humanos. Elaborar y proponer procedim. para la confec. de registros de datos estadísticos - Elaborar y proponer lineamientos para la elaborac.e implementac.de acciones. Diseñar herramientas de difusión sobre los ds de participación e información.Esta investigac. es de carácter descriptivo e interpretativo, analizará diferentes ftes de informac.con técnicas cuanti y cualitativas, durante período la promulgación de la Ley 26061/05 y la adhesión prov. 9396/07. Se tomarán del Sistema de Indicadores, el derecho a la educación, el tema transversal “Acceso a la información y participación”, y las categorías de análisis “Recepción del Derecho y Capacidades estatales”. Se trabajaran indicadores cuanti y señales de progreso (cualitativos): estructurales, proceso y resultados. Serán analizados los Programas implementados durante el período de estudio. Estrategias cuali observac. participante, entrevistas, grupos focales, talleres de sensibilización, etc .Otras ftes de info: doc. institucionales, registros, evaluac, contenidos de programas y campañas y de difusión, prensa escrita, legislaciones, etc. La info se analizará a través Programa Atlas Ti, y del análisis estadístico. Algunos resultados esperados: Evaluac.y ajuste del Sistema de indicadores, la sensibilizac. desde el EF de autorid. escol. para la formulac. de planes y programas para el monitoreo adecuado de los mismos y para la rendic. de ctas de su gestión. Estimular la participac. de NNA en la escuela. Realizac.de documental. Brindar herramientas al Edo para superv de la aplicación progresiva del ejercicio de los derechos de participación e información de NNA Promov la continua evaluac. de los programas y acc.desde el ED para estimular el reconoc. de la nec.de ajuste de la política. The issue of children from the rights focus implies a reading of the rights in which children and adolescents are the holders and the state actions that are implemented to enable the exercise of them. This perspective requires understanding the contents of the public policies, the participation level that is assigned, and the senses that they take in terms of rights progressivity of. The conception changes about children since the International Convention on the Rights of the Child and legislative changes challenge to the State and society to modify hegemony vision. Our hypothesis revolves around the following considerations: there have been some public policies and state actions changes, but they not turn out, yet, transformations in logical institutional and actors whose are encourages children participation in school. This could be reflects in the absence of qualitative analysis in the actions contents and the lack of sources specific statistics, that will be showing the control of the principal contents of integral protection. These, raise the questions that delimit our problem research: how is built citizenship exercise through children and adolescent’s participation in the school, and what are the institutional logical that empowers or turns difficult that exercise? And What are the institutional resources that make possible the effective exercise of the rights, through different programs and actions?. General Objective: To describe, analyze and interpret exercise progressivity participation and information rights that have the adolescents in secondary schools in Córdoba City during 2005 / 2010, in order to promote public spaces for the construction of citizenship. It will analyze three schools in Cordoba, provincial public school, national public school and private school. . Obj. Specifics: To analyze the formal and informal participative practices in the schools selected. - To apply quantitative and qualitative indicators of the “Indicators System for Infa

Differentiation of rat striatal embryonic stem cells in vitro: monolayer culture vs. three-dimensional coculture with differentiated brain cells.

Several groups have demonstrated the existence of self-renewing stem cells in embryonic and adult mouse brain. In vitro, these cells proliferate in response to epidermal growth factor, forming clusters of nestin-positive cells that may be dissociated and subcultured repetitively. Here we show that, in stem cell clusters derived from rat embryonic striatum, cell proliferation decreased with increasing number of passages and in response to elevated concentrations of potassium (30 mM KCl). In monolayer culture, the appearance of microtubule-associated protein type-5-immunoreactive (MAP-5(+)) cells (presumptive neurons) in response to basic fibroblast growth factor (bFGF) was reduced at low cell density and with increasing number of passages. In the presence of bFGF, elevated potassium caused a more differentiated neuronal phenotype, characterized by an increased proportion of MAP-5(+) cells, extensive neuritic branching, and higher specific activity of glutamic acid decarboxylase. Dissociated stem cells were able to invade cultured brain cell aggregates containing different proportions of neurons and glial cells, whereas they required the presence of a considerable proportion of glial cells in the host cultures to become neurofilament H-positive. The latter observation supports the view that astrocyte-derived factors influence early differentiation of the neuronal cell lineage.

Identification of cancer stem cells in Ewing's sarcoma

Summary : Cancer stem cells (CSC) that display tumor-initiating properties have recently been identified in several distinct types of malignancies, holding promise for more effective therapeutic strategies. However, evidence of such cells in sarcomas, which include some of the most aggressive and therapy-resistant tumors, has not been demonstrated to date. Here, we .identify and characterize cancer stem cells in Ewing's sarcoma family tumors (ESPY), a highly aggressive pediatric malignancy believed to be of mesenchymal stem cell (MSC) origin. Using magnetic bead cell separation of primary ESFT, we have isolated a subpopulation of CD133+ tumor cells that display the capacity to initiate and sustain tumor growth through serial transplantation in NOD/SCID mice, re-establishing at each in vivo passage the parental tumor phenotype and hierarchical cell organization. Consistent with the plasticity of MSCs, in vitro differentiation assays showed that the CD133+ cell population retained the ability to differentiate along adipogenic, osteogenic and chondrogenic lineages. Quantitative Real-Time PCR analysis of genes implicated in stem cell maintenance revealed that CD133+ ESFT cells express significantly higher levels of OCT4 and NANOG than their CD133- counterparts. Taken together, our observations provide the first identification of ESFT cancer stem cells (ET-CSC) and demonstration of their mesenchymal stem cell properties, a critical step toward a better biological understanding and rational therapeutic targeting of these tumors. Résumé : Des cellules souches tumorales avec des propriétés exclusives d'initiation tumorale ont récemment été identifiées dans différents types de cancers, permettant ainsi d'espérer le développement de thérapies plus efficaces. Cependant, l'existence de telles cellules dans les sarcomes, un sous-groupe de cancers d'origine mésenchymateuse très agressifs, n'a pas encore été démontrée. Dans ce travail de recherche, nous identifions et caractérisons des cellules souches tumorales dans le sarcome d'Ewing, une tumeur pédiatrique très agressive vraisemblablement dérivée de cellules souches mésenchymateuses (MSC). Afin de séparer des populations cellulaires dans des échantillons primaires de sarcome d'Ewing, nous avons utilisé des billes magnétiques couplées à des anticorps monoclonaux. Ceci nous a permis d'isoler une sous-population de cellules tumorales CD133+ qui ont la capacité d'initier et de maintenir la croissance tumorale dans des xénotransplantations en série effectuées dans des souris immunodéficientes NOD/SCID. Ces cellules reétablissent à chaque passage in vivo le phénotype de la tumeur d'origine ainsi que son organisation hiérarchique. En accord avec la plasticité des MSC, des tests de différentiation in vitro ont montré que les cellules CD133+ maintiennent la capacité de se différentier en adipocytes, ostéocytes et chondrocytes. Une analyse par PCR quantitative de gènes impliqués dans le maintien des cellules souches a montré que les cellules CD133+ expriment un niveau beaucoup plus élevé de OCT4 and NANOG que les cellules CD133-. En résumé, nos observations constituent la première identification de cellules souches tumorales dans le sarcome d'Ewing et démontrent leur propriété de cellules souches mésenchymateuses. Ceci constitue une étape clé vers une meilleure compréhension biologique et une meilleure approche thérapeutique de ces tumeurs.

Bcl9/Bcl9l are critical for Wnt-mediated regulation of stem cell traits in colon epithelium and adenocarcinomas.

Canonical Wnt signaling plays a critical role in stem cell maintenance in epithelial homeostasis and carcinogenesis. Here, we show that in the mouse this role is critically mediated by Bcl9/Bcl9l, the mammalian homologues of Legless, which in Drosophila is required for Armadillo/beta-catenin signaling. Conditional ablation of Bcl9/Bcl9l in the intestinal epithelium, where the essential role of Wnt signaling in epithelial homeostasis and stem cell maintenance is well documented, resulted in decreased expression of intestinal stem cell markers and impaired regeneration of ulcerated colon epithelium. Adenocarcinomas with aberrant Wnt signaling arose with similar incidence in wild-type and mutant mice. However, transcriptional profiles were vastly different: Whereas wild-type tumors displayed characteristics of epithelial-mesenchymal transition (EMT) and stem cell-like properties, these properties were largely abrogated in mutant tumors. These findings reveal an essential role for Bcl9/Bcl9l in regulating a subset of Wnt target genes involved in controlling EMT and stem cell-related features and suggest that targeting the Bcl9/Bcl9l arm of Wnt signaling in Wnt-activated cancers might attenuate these traits, which are associated with tumor invasion, metastasis, and resistance to therapy.

Cancer genes hypermethylated in human embryonic stem cells.

Developmental genes are silenced in embryonic stem cells by a bivalent histone-based chromatin mark. It has been proposed that this mark also confers a predisposition to aberrant DNA promoter hypermethylation of tumor suppressor genes (TSGs) in cancer. We report here that silencing of a significant proportion of these TSGs in human embryonic and adult stem cells is associated with promoter DNA hypermethylation. Our results indicate a role for DNA methylation in the control of gene expression in human stem cells and suggest that, for genes repressed by promoter hypermethylation in stem cells in vivo, the aberrant process in cancer could be understood as a defect in establishing an unmethylated promoter during differentiation, rather than as an anomalous process of de novo hypermethylation.

Accelerated telomere shortening in hematological lineages is limited to the first year following stem cell transplantation.

Using quantitative fluorescence in situ hybridization and flow cytometry, the telomere length of telomere repeat sequences after stem cell transplantation (SCT) were measured. The study included the telomeres of peripheral blood monocytes that should reflect the length of telomeres in stem cells and the telomeres of T lymphocytes that could shorten as a result of peripheral expansion. The loss of telomeres in monocytes and in memory T cells, although accelerated initially, became comparable to the loss of telomeres in healthy controls from the second year after transplantation. In addition, the telomere length in the naive T cells that were produced by the thymus was comparable to the telomere length in the naive T cells of the donor. Compared to the total length of telomeres available, the loss of telomere repeats in leukocytes after SCT resembles the accelerated shortening seen in early childhood and remains, therefore, relatively insignificant.

Identification of cancer stem cells in Ewing's sarcoma.

Cancer stem cells that display tumor-initiating properties have recently been identified in several distinct types of malignancies, holding promise for more effective therapeutic strategies. However, evidence of such cells in sarcomas, which include some of the most aggressive and therapy-resistant tumors, has not been shown to date. Here, we identify and characterize cancer stem cells in Ewing's sarcoma family tumors (ESFT), a highly aggressive pediatric malignancy believed to be of mesenchymal stem cell (MSC) origin. Using magnetic bead cell separation of primary ESFT, we have isolated a subpopulation of CD133+ tumor cells that display the capacity to initiate and sustain tumor growth through serial transplantation in nonobese diabetic/severe combined immunodeficiency mice, re-establishing at each in vivo passage the parental tumor phenotype and hierarchical cell organization. Consistent with the plasticity of MSCs, in vitro differentiation assays showed that the CD133+ cell population retained the ability to differentiate along adipogenic, osteogenic, and chondrogenic lineages. Quantitative real-time PCR analysis of genes implicated in stem cell maintenance revealed that CD133+ ESFT cells express significantly higher levels of OCT4 and NANOG than their CD133- counterparts. Taken together, our observations provide the first identification of ESFT cancer stem cells and demonstration of their MSC properties, a critical step towards a better biological understanding and rational therapeutic targeting of these tumors.

Decision-Making in Social Enterprises: Exploring the Link between Employee Participation and Organizational Commitment

This article studies the influence of the procedural justice resulting from participation in decision-making on employees' affective commitment in social enterprises. It also examines whether any potential link between participation and commitment is due to social exchange, as is the case with for-profit companies. The study is based on data from employees of French work integration social enterprises. The results confirm the positive relationship between procedural justice and affective commitment and the mediating role of perceived organizational support and leader-member exchanges. Managerial recommendations are then given to best maintain or increase employees' involvement in the decision-making processes of social enterprises.