Co-administration of human MSC overexpressing HIF-1α increases human CD34+ cell engraftment in vivo

Background: Poor graft function or graft failure after allogeneic stem cell transplantation is an unmet medical need, in which mesenchymal stromal cells (MSC) constitute an attractive potential therapeutic approach. Hypoxia-inducible factor-1α (HIF-1α) overexpression in MSC (HIF-MSC) potentiates the angiogenic and immunomodulatory properties of these cells, so we hypothesized that co-transplantation of MSC-HIF with CD34+ human cord blood cells would also enhance hematopoietic stem cell engraftment and function both in vitro and in vivo. Methods: Human MSC were obtained from dental pulp. Lentiviral overexpression of HIF-1α was performed transducing cells with pWPI-green fluorescent protein (GFP) (MSC WT) or pWPI-HIF-1α-GFP (HIF-MSC) expression vectors. Human cord blood CD34+ cells were co-cultured with MSC WT or HIF-MSC (4:1) for 72 h. Then, viability (Annexin V and 7-AAD), cell cycle, ROS expression and immunophenotyping of key molecules involved in engraftment (CXCR4, CD34, ITGA4, c-KIT) were evaluated by flow cytometry in CD34+ cells. In addition, CD34+ cells clonal expansion was analyzed by clonogenic assays. Finally, in vivo engraftment was measured by flow cytometry 4-weeks after CD34+ cell transplantation with or without intrabone MSC WT or HIF-MSC in NOD/SCID mice. Results: We did not observe significant differences in viability, cell cycle and ROS expression between CD34+ cells co-cultured with MSC WT or HIF-MSC. Nevertheless, a significant increase in CD34, CXCR4 and ITGA4 expression (p = 0.009; p = 0.001; p = 0.013, respectively) was observed in CD34+ cells co-cultured with HIF-MSC compared to MSC WT. In addition, CD34+ cells cultured with HIF-MSC displayed a higher CFU-GM clonogenic potential than those cultured with MSC WT (p = 0.048). We also observed a significant increase in CD34+ cells engraftment ability when they were co-transplanted with HIF-MSC compared to CD34+ co-transplanted with MSC WT (p = 0.016) or alone (p = 0.015) in both the injected and contralateral femurs (p = 0.024, p = 0.008 respectively). Conclusions: Co-transplantation of human CD34+ cells with HIF-MSC enhances cell engraftment in vivo. This is probably due to the ability of HIF-MSC to increase clonogenic capacity of hematopoietic cells and to induce the expression of adhesion molecules involved in graft survival in the hematopoietic niche

Challenges of the use of new generation massive sequencing (NGS) of the benthic macrofauna for the evaluation of the marine environment quality.

La Directiva Marco del Agua 2000/60/CE obliga al diagnóstico ambiental del ecosistema marino, incluyendo la evaluación de las especies de macroinvertebrados considerados bioindicadores presentes en el medio. Hasta la fecha, este tipo de determinaciones se realizan mediante la identificación taxonómica de visu de la macrofauna bentónica presente en las muestras y el cálculo de bioíndices asociados, un proceso costoso en términos de tiempo y financiación y, en algunos casos, subjetivo por precisar de un equipo humano altamente especializado y por la dificultad de identificar correctamente determinadas especies. En este sentido, las técnicas de DNA barcoding permiten identificar de forma fiable organismos empleando técnicas de secuenciación de DNA y evitando las desventajas de la identificación morfotaxonómica. Por otro lado, el reciente desarrollo de técnicas de secuenciación masiva de DNA de nueva generación (NGS) ha permitido el desarrollo del DNA metabarcoding, o caracterización de poblaciones de organismos presentes en una muestra empleando datos genómicos. Este trabajo plantea los retos fundamentales que presenta, a día de hoy, el análisis de organismos bioindicadores de calidad ambiental marina a través de las técnicas de secuenciación NGS.The Water Framework Directive 2000/60/EC regulates the environmental diagnosis of the marine ecosystem, including the evaluation of species of bioindicator macroinvertebrates present in the environment. To date, these types of determinations are carried out through the morphotaxonomic identification of the benthic macrofauna present in the samples and the calculation of associated biotic indexes, a process that is time-consuming and resource-intensive, being in some cases inaccurate due to the requirement of highly specialized human resources and the difficulty of correctly identifying certain species. In this respect, DNA barcoding techniques allow the reliable identification of organisms using DNA sequencing techniques and avoiding the disadvantages of morphotaxonomic identification. On the other hand, the recent development of New Generation DNA Sequencing techniques (NGS) has allowed the development of DNA metabarcoding, i.e. the characterization of populations of organisms present in a sample using genomic data. This paper shows the fundamental challenges to be overcome in order to establish a NGS sequencing-based assessment of the marine environmental quality.Ciencias Experimentale

Retos del empleo de la secuenciación masiva de nueva generación (NGS) de la macrofauna bentónica para la evaluación de la calidad ambiental marina

The Water Framework Directive 2000/60/EC regulates the environmental diagnosis of the marine ecosystem, including the evaluation of species of bioindicator macroinvertebrates present in the environment. To date, these types of determinations are carried out through the morphotaxonomic identification of the benthic macrofauna present in the samples and the calculation of associated biotic indexes, a process that is time-consuming and resource-intensive, being in some cases inaccurate due to the requirement of highly specialized human resources and the difficulty of correctly identifying certain species. In this respect, DNA barcoding techniques allow the reliable identification of organisms using DNA sequencing techniques and avoiding the disadvantages of morphotaxonomic identification. On the other hand, the recent development of New Generation DNA Sequencing techniques (NGS) has allowed the development of DNA metabarcoding, i.e. the characterization of populations of organisms present in a sample using genomic data. This paper shows the fundamental challenges to be overcome in order to establish a NGS sequencing-based assessment of the marine environmental quality.La Directiva Marco del Agua 2000/60/CE obliga al diagnóstico ambiental del ecosistema marino, incluyendo la evaluación de las especies de macroinvertebrados considerados bioindicadores presentes en el medio. Hasta la fecha, este tipo de determinaciones se realizan mediante la identificación taxonómica de visu de la macrofauna bentónica presente en las muestras y el cálculo de bioíndices asociados, un proceso costoso en términos de tiempo y financiación y, en algunos casos, subjetivo por precisar de un equipo humano altamente especializado y por la dificultad de identificar correctamente determinadas especies. En este sentido, las técnicas de DNA barcoding permiten identificar de forma fiable organismos empleando técnicas de secuenciación de DNA y evitando las desventajas de la identificación morfotaxonómica. Por otro lado, el reciente desarrollo de técnicas de secuenciación masiva de DNA de nueva generación (NGS) ha permitido el desarrollo del DNA metabarcoding, o caracterización de poblaciones de organismos presentes en una muestra empleando datos genómicos. Este trabajo plantea los retos fundamentales que presenta, a día de hoy, el análisis de organismos bioindicadores de calidad ambiental marina a través de las técnicas de secuenciación NGS

Diet quality index as a predictor of treatment efficacy in overweight and obese adolescents: The EVASYON study

The EVASYON Study Group.[Background & aim]: A diet quality index (DQI) is a tool that provides an overall score of an individual's dietary intake when assessing compliance with food-based dietary guidelines. A number of DQIs have emerged, albeit their associations with health-related outcomes are debated. The aim of the present study was to assess whether adherence to dietary intervention, and the overall quality of the diet, can predict body composition changes.[Methods]: To this purpose, overweight/obese adolescents (n = 117, aged: 13–16 years; 51 males, 66 females) were recruited into a multi-component (diet, physical activity and psychological support) family-based group treatment programme. We measured the adolescents' compliance and body composition at baseline and after 2 months (intensive phase) and 13 months (extensive phase) of follow-up. Also, at baseline, after 6 months, and at the end of follow-up we calculated the DQI.[Results]: Global compliance with the dietary intervention was 37.4% during the intensive phase, and 14.3% during the extensive phase. Physical activity compliance was 94.1% at 2-months and 34.7% at 13months and psychological support compliance were growing over the intervention period (10.3% intensive phase and 45.3% during extensive phase). Adolescents complying with the meal frequency criteria at the end of the extensive phase had greater reductions in FMI z-scores than those did not complying (Cohen's d = 0.53). A statistically significant association was observed with the diet quality index. DQI-A variation explained 98.1% of BMI z-score changes and 95.1% of FMI changes.[Conclusions]: We conclude that assessment of changes in diet quality could be a useful tool in predicting body composition changes in obese adolescents involved in a diet and physical activity intervention programme backed-up by psychological and family support.The study was supported by the Ministry of Health, Social Services and Equality via the Carlos III Institute of Health (FIS Grant PI051080, PI051579). The EVASYON study received the award for the best applied research project in 2009 from AESAN (Spanish Agency for Food Safety and Nutrition from the Spanish Ministry of Health and Consumer Affairs. The study was supported by Aragon's Regional Government (DGA, Diputación General de Aragón) and European Regional Development Fund.Peer reviewe