Combined multidimensional single-cell protein and RNA profiling dissects the cellular and functional heterogeneity of thymic epithelial cells.

The network of thymic stromal cells provides essential niches with unique molecular cues controlling T cell development and selection. Recent single-cell RNA sequencing studies have uncovered previously unappreciated transcriptional heterogeneity among thymic epithelial cells (TEC). However, there are only very few cell markers that allow a comparable phenotypic identification of TEC. Here, using massively parallel flow cytometry and machine learning, we deconvoluted known TEC phenotypes into novel subpopulations. Using CITEseq, these phenotypes were related to corresponding TEC subtypes defined by the cells' RNA profiles. This approach allowed the phenotypic identification of perinatal cTEC and their physical localisation within the cortical stromal scaffold. In addition, we demonstrate the dynamic change in the frequency of perinatal cTEC in response to developing thymocytes and reveal their exceptional efficiency in positive selection. Collectively, our study identifies markers that allow for an unprecedented dissection of the thymus stromal complexity, as well as physical isolation of TEC populations and assignment of specific functions to individual TEC subtypes.

Ageing compromises mouse thymus function and remodels epithelial cell differentiation.

Ageing is characterised by cellular senescence, leading to imbalanced tissue maintenance, cell death and compromised organ function. This is first observed in the thymus, the primary lymphoid organ that generates and selects T cells. However, the molecular and cellular mechanisms underpinning these ageing processes remain unclear. Here, we show that mouse ageing leads to less efficient T cell selection, decreased self-antigen representation and increased T cell receptor repertoire diversity. Using a combination of single-cell RNA-seq and lineage-tracing, we find that progenitor cells are the principal targets of ageing, whereas the function of individual mature thymic epithelial cells is compromised only modestly. Specifically, an early-life precursor cell population, retained in the mouse cortex postnatally, is virtually extinguished at puberty. Concomitantly, a medullary precursor cell quiesces, thereby impairing maintenance of the medullary epithelium. Thus, ageing disrupts thymic progenitor differentiation and impairs the core immunological functions of the thymus.

Prescripción enfermera: análisis de concepto / Nurse prescription: A concept analysis

Objetivo: Examinar críticamente el concepto de prescripción enfermera mediante el estudio de los antecedentes y una revisión de la literatura científica con el fin de desarrollar una concepción precisa de esta actuación enfermera e identificar los elementos esenciales que envuelven este concepto. Método: Aplicación del procedimiento de análisis de concepto descrito por Wilson y adaptado por Avant. Resultados: El concepto de prescripción enfermera implica prescribir por parte de la enfermera el mejor régimen terapéutico frente a un problema de salud. Esta prescripción estará guiada por la valoración del problema de salud, por el criterio de la buena práctica clínica del profesional enfermero y estará dirigida a satisfacer las necesidades de salud del usuario y la población. Conclusiones: Los resultados aclaran el significado del concepto de estudio para ayudar a los profesionales a comprender y abordar esta actuación en todas sus dimensiones y promover el reconocimiento social de la profesión enfermera

Objective: To critically analyse the concept of nursing prescription through the study of its background and a review of the scientific literature, in order to develop an accurate conception of this nursing activity and to identify the essential elements surrounding this concept. Method: Application of the concept analysis method described by Wilson, and adapted by Avant. Results: The concept of nurse prescription implies prescribing, by the nurse, the best therapeutic regimen for a health problem. This prescription will be guided by the assessment of the health problem, by the criterion of the good clinical practice of the nurse, and will be focused to satisfy the health needs of the patient and the population. Conclusions: The results clarify the meaning of the study's concept to help professionals understand and address this nursing activity in all its dimensions, and promote social recognition of the nursing profession

[Nurse prescription: A concept analysis]. / Prescripción enfermera: análisis de concepto.

OBJECTIVE: To critically analyse the concept of nursing prescription through the study of its background and a review of the scientific literature, in order to develop an accurate conception of this nursing activity and to identify the essential elements surrounding this concept. METHOD: Application of the concept analysis method described by Wilson, and adapted by Avant. RESULTS: The concept of nurse prescription implies prescribing, by the nurse, the best therapeutic regimen for a health problem. This prescription will be guided by the assessment of the health problem, by the criterion of the good clinical practice of the nurse, and will be focused to satisfy the health needs of the patient and the population. CONCLUSIONS: The results clarify the meaning of the study's concept to help professionals understand and address this nursing activity in all its dimensions, and promote social recognition of the nursing profession.

DN2 Thymocytes Activate a Specific Robust DNA Damage Response to Ionizing Radiation-Induced DNA Double-Strand Breaks.

For successful bone marrow transplantation (BMT), a preconditioning regime involving chemo and radiotherapy is used that results in DNA damage to both hematopoietic and stromal elements. Following radiation exposure, it is well recognized that a single wave of host-derived thymocytes reconstitutes the irradiated thymus, with donor-derived thymocytes appearing about 7 days post BMT. Our previous studies have demonstrated that, in the presence of donor hematopoietic cells lacking T lineage potential, these host-derived thymocytes are able to generate a polyclonal cohort of functionally mature peripheral T cells numerically comprising ~25% of the peripheral T cell pool of euthymic mice. Importantly, we demonstrated that radioresistant CD44+ CD25+ CD117+ DN2 progenitors were responsible for this thymic auto-reconstitution. Until recently, the mechanisms underlying the radioresistance of DN2 progenitors were unknown. Herein, we have used the in vitro "Plastic Thymus" culture system to perform a detailed investigation of the mechanisms responsible for the high radioresistance of DN2 cells compared with radiosensitive hematopoietic stem cells. Our results indicate that several aspects of DN2 biology, such as (i) rapid DNA damage response (DDR) activation in response to ionizing radiation-induced DNA damage, (ii) efficient repair of DNA double-strand breaks, and (iii) induction of a protective G1/S checkpoint contribute to promoting DN2 cell survival post-irradiation. We have previously shown that hypoxia increases the radioresistance of bone marrow stromal cells in vitro, at least in part by enhancing their DNA double-strand break (DNA DSB) repair capacity. Since the thymus is also a hypoxic environment, we investigated the potential effects of hypoxia on the DDR of DN2 thymocytes. Finally, we demonstrate for the first time that de novo DN2 thymocytes are able to rapidly repair DNA DSBs following thymic irradiation in vivo.

The Transcription Factor Hif-1 Enhances the Radio-Resistance of Mouse MSCs.

Mesenchymal stromal cells (MSCs) are multipotent progenitors supporting bone marrow hematopoiesis. MSCs have an efficient DNA damage response (DDR) and are consequently relatively radio-resistant cells. Therefore, MSCs are key to hematopoietic reconstitution following total body irradiation (TBI) and bone marrow transplantation (BMT). The bone marrow niche is hypoxic and via the heterodimeric transcription factor Hypoxia-inducible factor-1 (Hif-1), hypoxia enhances the DDR. Using gene knock-down, we have previously shown that the Hif-1α subunit of Hif-1 is involved in mouse MSC radio-resistance, however its exact mechanism of action remains unknown. In order to dissect the involvement of Hif-1α in the DDR, we used CRISPR/Cas9 technology to generate a stable mutant of the mouse MSC cell line MS5 lacking Hif-1α expression. Herein, we show that it is the whole Hif-1 transcription factor, and not only the Hif-1α subunit, that modulates the DDR of mouse MSCs. This effect is dependent upon the presence of a Hif-1α protein capable of binding to both DNA and its heterodimeric partner Arnt (Hif-1ß). Detailed transcriptomic and proteomic analysis of Hif1a KO MS5 cells leads us to conclude that Hif-1α may be acting indirectly on the DNA repair process. These findings have important implications for the modulation of MSC radio-resistance in the context of BMT and cancer.

Implantación de la gestión enfermera de la demanda en las entidades proveedoras de servicios de Atención Primaria de Salud de Cataluña / Implementation of nurse demand managment in primary health care service providers in Catalonia

Objetivo: Describir y analizar la aplicación de la gestión enfermera de la demanda (GED) en los distintos proveedores de salud de Atención Primaria de Salud en Cataluña desde 2005 hasta finales de 2014. Diseño: Estudio transversal. Participantes: Todos los proveedores de servicios de Atención Primaria de Salud en Cataluña (n = 37). Mediciones principales: Encuestas con la dirección de enfermería de cada proveedor sobre las dificultades y elementos facilitadores en relación a la implantación de la GED. Los factores facilitadores y las barreras se clasificaron en 3 tipos: a) profesionales de la salud (competencias, actitudes, motivación para el cambio y características individuales); b) contexto social (pacientes y acompañantes), y c) factores relacionados con el sistema (organización y estructura, incentivos económicos). Resultados: De los 37 proveedores, 26 (70,3%) han aplicado la GED. Las principales barreras identificadas son la regulación de la prescripción enfermera, la falta de conocimientos y habilidades de las enfermeras y la falta de protocolos en el inicio de la implantación. Entre los factores facilitadores destacan la formación específica de los profesionales, una mayor proporción de ratio de enfermeras por cada médico (médico de familia o pediatra), consenso en los circuitos con todos los profesionales y la vinculación de la introducción de la GED a incentivos económicos. Conclusiones: La GED se está consolidando en Cataluña. Sin embargo, la GED debería ser introducida en el plan de estudios de grado de enfermería y programas de educación continua en equipos de atención primaria (AU)

Objective: To describe and analyse the implementaction of nurse demand managment (NDM) mong health care providers in Catalonia from 2005 to 2014. Design: Cross sectional survey. Participants All service providers in Catalonia (n=37). Main measurements Interviews with nurse manager of each health care provides about ht barriers and facilitators concerning NDM. Facilitators and barriers were classified into 3 types: (I) health professional (competence, attitudes, motivation for change and individual characteristics); (II) social context (patients and companions), and (III) system related factors (organization and structure, economic incentives). Results: Of the 37 providers, 26 (70.3%) have implemented the Demand Management Nurse (NDM). The main barriers identified are the nurse prescriptin regulation, lack of knowledge and skills of nurses, and the lack of protocols at the start of implantation. Among the facilitators are the specific training of professionals, a higher ratio of nurses to doctors, consensus circuits with all professionals and linking the implementation of NDM to economic incentives. Discussion: NDM is consolidated in Catalonia. However, the NDM should be included in the curricula of nursing degree and continuing education programs in primary care teams (AU)

Differential Response of Mouse Thymic Epithelial Cell Types to Ionizing Radiation-Induced DNA Damage.

Thymic epithelial cells (TECs) are the main components of the thymic stroma that support and control T-cell development. Preparative regimens using DNA-damaging agents, such as total body irradiation and/or chemotherapeutic drugs, that are necessary prior to bone marrow transplantation (BMT) have profound deleterious effects on the hematopoietic system, including the thymic stroma, which may be one of the main causes for the prolonged periods of T-cell deficiency and the inefficient T cell reconstitution that are common following BMT. The DNA damage response (DDR) is a complex signaling network that allows cells to respond to all sorts of genotoxic insults. Hypoxia is known to modulate the DDR and play a role affecting the survival capacity of different cell types. In this study, we have characterized in detail the DDR of cortical and medullary TEC lines and their response to ionizing radiation, as well as the effects of hypoxia on their DDR. Although both mTECs and cTECs display relatively high radio-resistance, mTEC cells have an increased survival capacity to ionizing radiation (IR)-induced DNA damage, and hypoxia specifically decreases the radio-resistance of mTECs by upregulating the expression of the pro-apoptotic factor Bim. Analysis of the expression of TEC functional factors by primary mouse TECs showed a marked decrease of highly important genes for TEC function and confirmed cTECs as the most affected cell type by IR. These findings have important implications for improving the outcomes of BMT and promoting successful T cell reconstitution.

[Implementation of nurse demand managment in primary health care service providers in Catalonia]. / Implantación de la gestión enfermera de la demanda en las entidades proveedoras de servicios de Atención Primaria de Salud de Cataluña.

OBJECTIVE: To describe and analyse the implementaction of nurse demand managment (NDM) among health care providers in Catalonia from 2005 to 2014. DESIGN: Cross sectional survey. Participants All service providers in Catalonia (n=37). Main measurements Interviews with nurse manager of each health care provides about ht barriers and facilitators concerning NDM. Facilitators and barriers were classified into 3 types: (i)health professional (competence, attitudes, motivation for change and individual characteristics); (ii)social context (patients and companions), and (iii)system related factors (organization and structure, economic incentives). RESULTS: Of the 37 providers, 26 (70.3%) have implemented the Demand Management Nurse (NDM). The main barriers identified are the nurse prescriptin regulation, lack of knowledge and skills of nurses, and the lack of protocols at the start of implantation. Among the facilitators are the specific training of professionals, a higher ratio of nurses to doctors, consensus circuits with all professionals and linking the implementation of NDM to economic incentives. DISCUSSION: NDM is consolidated in Catalonia. However, the NDM should be included in the curricula of nursing degree and continuing education programs in primary care teams.

Importance of Ecological Factors and Colony Handling for Optimizing Health Status of Apiaries in Mediterranean Ecosystems.

We analyzed six apiaries in several natural environments with a Mediterranean ecosystem in Madrid, central Spain, in order to understand how landscape and management characteristics may influence apiary health and bee production in the long term. We focused on five criteria (habitat quality, landscape heterogeneity, climate, management and health), as well as 30 subcriteria, and we used the analytic hierarchy process (AHP) to rank them according to relevance. Habitat quality proved to have the highest relevance, followed by beehive management. Within habitat quality, the following subcriteria proved to be most relevant: orographic diversity, elevation range and important plant species located 1.5 km from the apiary. The most important subcriteria under beehive management were honey production, movement of the apiary to a location with a higher altitude and wax renewal. Temperature was the most important subcriterion under climate, while pathogen and Varroa loads were the most significant under health. Two of the six apiaries showed the best values in the AHP analysis and showed annual honey production of 70 and 28 kg/colony. This high productivity was due primarily to high elevation range and high orographic diversity, which favored high habitat quality. In addition, one of these apiaries showed the best value for beehive management, while the other showed the best value for health, reflected in the low pathogen load and low average number of viruses. These results highlight the importance of environmental factors and good sanitary practices to maximize apiary health and honey productivity.

Vigor for in vitro culture traits in S. melongena × S. aethiopicum hybrids with potential as rootstocks for eggplant.

Hybrids of Solanum melongena and S. aethiopicum are of interest as rootstocks of eggplant, as they are highly vigorous and can incorporate resistance to several diseases. However, hybridization between both species is difficult. Therefore, protocols for in vitro culture are of great interest for their micropropagation and biotechnological breeding. We assessed the organogenesis response from leaf explants in four interspecific hybrids and in their parents testing two organogenic media: SIM-A, containing 6-benzylaminopurine and kinetin, and SIM-B, which contains thidiazuron. A higher regeneration capacity in the hybrids compared to their parents was observed. Whereas in interspecific hybrids and in one accession of S. melongena similar regeneration rates were observed for SIM-A and SIM-B, higher regeneration was found in the rest of genotypes when thidiazuron was used. Rooting ability in the interspecific hybrids was lower in in vitro micropropagated plants (35-60%) than in plants regenerated from explants (100%). The addition of indolbutiric acid (1 mg L(-1)) induced roots in nonrooted genotypes. In summary, we have adjusted in vitro culture conditions for regenerating and rooting S. melongena × S. aethiopicum hybrids. We have also demonstrated that these hybrids are heterotic for regeneration, which may be of interest for basic science studies.

ATR activates the S-M checkpoint during unperturbed growth to ensure sufficient replication prior to mitotic onset.

Cells must accurately replicate and segregate their DNA once per cell cycle in order to successfully transmit genetic information. During S phase in the presence of agents that cause replication stress, ATR-dependent checkpoints regulate origin firing and the replication machinery as well as prevent untimely mitosis. Here, we investigate the role of ATR during unperturbed growth in vertebrate cells. In the absence of ATR, individual replication forks progress more slowly, and an increased number of replication origins are activated. These cells also enter mitosis early and divide more rapidly, culminating in chromosome bridges and laggards at anaphase, failed cytokinesis, and cell death. Interestingly, cell death can be rescued by prolonging mitosis with partial inhibition of the mitotic cyclin-dependent kinase 1. Our data indicate that one of the essential roles of ATR during normal growth is to minimize the level of unreplicated DNA before the onset of mitosis.