Extracellular vesicle and soluble fractions of adipose tissue-derived mesenchymal stem cells secretome induce inflammatory cytokines modulation in an in vitro model of discogenic pain

.BACKGROUND CONTEXT: Mesenchymal Stem Cells (MSCs) secretome or conditioned medium (CM) is a complex cocktail of different molecules, some of which, particularly those con- tained in extracellular vesicles, already have proven therapeutic applications. PURPOSE: CM may well represent promising therapy for discogenic pain and the intention of this work is to assess its therapeutic potential using an in vitro model of this condition. STUDY DESIGN: This is an experimental study. METHODS: Our in vitro model comprised Nucleus Pulposus (NP) and Annulus Fibrosus (AF) cells inflamed with TNF. To assess the potential therapeutic value of CM and its components, extracellular vesicles (EVs) and soluble culture fraction (SF), cell inflammation took place under 3 different conditions: either in the presence of whole CM, isolated EVs or SF, and concentrations of pro-inflammatory cytokines, metalloproteinases (MMPs) and neurotrophic factors produced in all 3 cases were compared. RESULTS: In the presence of whole CM, both in vitro gene expression by the NP and AF test cells and analysis of their protein content showed high modulatory effects on inflammation and MMP inhibition. The presence of EVs and SF showed similar but much smaller effects, and this was par- ticularly marked in the case of NP cells. CONCLUSIONS: Our results show that, compared to EVs and SF, the presence of whole CM has the greatest positive effect on the modulation of pro-inflammatory and catabolic factors. These observations suggest that CM could protect against inflammation and the resulting intervertebral disc (IVD) degeneration that leads to discogenic pain. CLINICAL SIGNIFICANCE: Many patients' expectations are not met by current non-operative and surgical treatments for discogenic low back pain. We propose the use of the MSCs secretomeS

The follower location problem with attraction thresholds

Abstract. In this research note, the follower location problem where the customers' choice is based on an attraction function is analysed. The attraction function depends on both the distance between customers and facilities, and the characteristics (quality) of the facilities. Customers at each node impose a minimum level of attraction in order to patronise a facility and then they share their buying power among the facilities that pass this threshold. The amount of demand captured by each of these facilities is proportional to the attraction perceived by the customers. In this case, a discretisation of this network problem is proved. JEL classification: L10, R3

Spatial competition in networks under delivered pricing *

Abstract. We consider a two-stage non-cooperative Bertrand game with location choice involving r firms. There are n spatially separated markets located at the vertices of a network. Each firm first selects the location of a facility and then selects the delivered price in the markets in order to maximise its profit. The article extends the duopolistic model with completely inelastic demand (Lederer and Thisse 1990) to the oligopolistic scenario. Under moderate assumptions, a pure strategy equilibrium, which minimises social costs, exists. Furthermore, an equilibrium location can be obtained by finite steps and consists of vertices only. JEL classification: L13, R32, D4

Marco teórico y metodológico de educación ambiental e intercultural para un desarrollo sostenible

El actual modelo socioeconómico de las sociedades de los países desarrollados es insostenible, pues lleva aparejados importantes problemas ambientales que ponen en riesgo la supervivencia de la Tierra, por lo que es necesario un nuevo modelo de sociedad que haga posible un futuro más sostenible y, en ello, la educación juega un papel clave. Se presenta una propuesta educativa que pretende integrar la perspectiva constructivista, concretamente el aprendizaje por investigación; pero, además, intenta mostrar la “aplicabilidad” de la Educación Ambiental e Intercultural para un Desarrollo Sostenible a la resolución de problemas reales y concretos, que salvando la distancia entre la teoría y la práctica, consiga la transformación de las actitudes, conocimientos y comportamientos de la comunidad educativa a nivel local y global. Además, introduce los contenidos de la gestión sostenible en el currículo y permite unir la práctica y la enseñanza de la sostenibilidad.Palabras clave: Educación Ambiental; Educación Intercultural; Desarrollo Sostenible; Propuesta metodológica.Theoretical and methodological frame of environmental and intercultural education for a sustainable developmentThe current model of society in developed countries is unbearable because it carries with it main environmental problems that put in risk the survival of the Earth, so that, a new model of society that makes a more sustainable future possible is necessary. In doing so, the education plays a key role. We present an educational proposal that tries to integrate the constructivist perspective, more precisely the learning by research methodology; but also tries to show that Environmental and Intercultural Education for a Sustainable Development can be applied to the solving of real problems, saving the gap between theory and practice to achieve the change of attitudes, knowledge and behaviours of the educational community in a more local and global level. Also, it introduces the content of the sustainable management of resources in the curriculum and makes it possible to put together the practice and teaching of sustainability.Key words: Environmental Education; Intercultural Education; Sustainable Development; Methodological proposal

Gene Networks Driving Genetic Variation in Milk and Cheese-Making Traits of Spanish Assaf Sheep

Artículo de InvestigaciónMost of the milk produced by sheep is used for the production of high-quality cheese. Consequently, traits related to milk coagulation properties and cheese yield are economically important to the Spanish dairy industry. The present study aims to identify candidate genes and their regulators related to 14 milk and cheese-making traits and to develop a low-density panel of markers that could be used to predict an individual’s genetic potential for cheese-making efficiency. In this study, we performed a combination of the classical genome-wide association study (GWAS) with a stepwise regression method and a pleiotropy analysis to determine the best combination of the variants located within the confidence intervals of the potential candidate genes that may explain the greatest genetic variance for milk and cheese-making traits. Two gene networks related to milk and cheese-making traits were created using the genomic relationship matrices built through a stepwise multiple regression approach. Several co-associated genes in these networks are involved in biological processes previously found to be associated with milk synthesis and cheese-making efficiency. The methodology applied in this study enabled the selection of a co-association network comprised of 374 variants located in the surrounding of genes showing a potential influence on milk synthesis and cheese-making efficiency.S

Genome-wide association studies (GWAS) and post-GWAS analyses for technological traits in Assaf and Churra dairy breeds

20 p.This study aimed to perform a GWAS to identify genomic regions associated with milk and cheese-making traits in Assaf and Churra dairy sheep breeds; second, it aimed to identify possible positional and functional candidate genes and their interactions through post-GWAS studies. For 2,020 dairy ewes from 2 breeds (1,039 Spanish Assaf and 981 Churra), milk samples were collected and analyzed to determine 6 milk production and composition traits and 6 traits related to milk coagulation properties and cheese yield. The genetic profiles of the ewes were obtained using a genotyping chip array that included 50,934 SNP markers. For both milk and cheese-making traits, separate single-breed GWAS were performed using GCTA software. The set of positional candidate genes identified via GWAS was subjected to guilt-by-association-based prioritization analysis with ToppGene software. Totals of 84 and 139 chromosome-wise significant associations for the 6 milk traits and the 6 cheese-making traits were identified in this study. No significant SNPs were found in common between the 2 studied breeds, possibly due to their genetic heterogeneity of the phenotypes under study. Additionally, 63 and 176 positional candidate genes were located in the genomic intervals defined as confidence regions in relation to the significant SNPs identified for the analyzed traits for Assaf and Churra breeds. After the functional prioritization analysis, 71 genes were identified as promising positional and functional candidate genes and proposed as targets of future research to identify putative causative variants in relation to the traits under examination. In addition, this multitrait study allowed us to identify variants that have a pleiotropic effect on both milk production and cheese-related traits. The incorporation of variants among the proposed functional and positional candidate genes into genomic selection strategies represent an interesting approach for achieving rapid genetic gains, specifically for those traits difficult to measure, such as cheese-making traits.S

Differences Between High vs. Low Performance Chess Players in Heart Rate Variability During Chess Problems

Background: Heart rate variability (HRV) has been considered as a measure of heart-brain interaction and autonomic modulation, and it is modified by cognitive and attentional tasks. In cognitive tasks, HRV was reduced in participants who achieved worse results. This could indicate the possibility of HRV predicting cognitive performance, but this association is still unclear in a high cognitive load sport such as chess.Objective: To analyze modifications on HRV and subjective perception of stress, difficulty and complexity in different chess problem tasks.Design: HRV was assessed at baseline. During the chess problems, HRV was also monitored, and immediately after chess problems the subjective stress, difficulty and complexity were also registered.Methods: A total of 16 male chess players, age: 35.19 (13.44) and ELO: 1927.69 (167.78) were analyzed while six chess problem solving tasks with different level of difficulty were conducted (two low level, two medium level and two high level chess problems). Participants were classified according to their results into two groups: high performance or low performance.Results: Friedman test showed a significant effect of tasks in HRV indexes and perceived difficulty, stress and complexity in both high and low performance groups. A decrease in HRV was observed in both groups when chess problems difficulty increased. In addition, HRV was significantly higher in the high performance group than in the low performance group during chess problems.Conclusion: An increase in autonomic modulation was observed to meet the cognitive demands of the problems, being higher while the difficulty of the tasks increased. Non-linear HRV indexes seem to be more reactive to tasks difficulty, being an interesting and useful tool in chess training

Introgression and pyramiding into common bean market class fabada of genes conferring resistance to anthracnose and potyvirus

Anthracnose and bean common mosaic (BCM) are considered major diseases in common bean crop causing severe yield losses worldwide. This work describes the introgression and pyramiding of genes conferring genetic resistance to BCM and anthracnose local races into line A25, a bean genotype classified as market class fabada. Resistant plants were selected using resistance tests or combining resistance tests and marker-assisted selection. Lines A252, A321, A493, Sanilac BC6-Are, and BRB130 were used as resistance sources. Resistance genes to anthracnose (Co-2 ( C ), Co-2 ( A252 ) and Co-3/9) and/or BCM (I and bc-3) were introgressed in line A25 through six parallel backcrossing programs, and six breeding lines showing a fabada seed phenotype were obtained after six backcross generations: line A1258 from A252; A1231 from A321; A1220 from A493; A1183 and A1878 from Sanilac BC6-Are; and line A2418 from BRB130. Pyramiding of different genes were developed using the pedigree method from a single cross between lines obtained in the introgression step: line A1699 (derived from cross A1258 × A1220), A2438 (A1220 × A1183), A2806 (A1878 × A2418), and A3308 (A1699 × A2806). A characterization based on eight morpho-agronomic traits revealed a limited differentiation among the obtained breeding lines and the recurrent line A25. However, using a set of seven molecular markers linked to the loci used in the breeding programs it was possible to differentiate the 11 fabada lines. Considering the genetic control of the resistance in resistant donor lines, the observed segregations in the last backcrossing generation, the reaction against the pathogens, and the expression of the molecular markers it was also possible to infer the genotype conferring resistance in the ten fabada breeding lines obtained. As a result of these breeding programs, genetic resistance to three anthracnose races controlled by genes included in clusters Co-2 and Co-3/9, and genetic resistance to BCM controlled by genotype I + bc-3 was combined in the fabada line A3308

c-Jun N-Terminal Kinase 1 Is Required for Toll-Like Receptor 1 Gene Expression in Macrophages

P. 5027-5034The regulation of innate immune responses to pathogens occurs through the interaction of Toll-like receptors (TLRs) with pathogen-associated molecular patterns and the activation of several signaling pathways whose contribution to the overall innate immune response to pathogens is poorly understood. We demonstrate a mechanism of control of murine macrophage responses mediated by TLR1/2 heterodimers through c-Jun N-terminal kinase 1 (JNK1) activity. JNK controls tumor necrosis factor alpha production and TLR-mediated macrophage responses to Borrelia burgdorferi, the causative agent of Lyme disease, and the TLR1/TLR2-specific agonist PAM3CSK4. JNK1, but not JNK2, activity regulates the expression of the tlr1 gene in the macrophage cell line RAW264.7, as well as in primary CD11b cells. We also show that the proximal promoter region of the human tlr1 gene contains an AP-1 binding site that is subjected to regulation by the kinase and binds two complexes that involve the JNK substrates c-Jun, JunD, and ATF-2. These results demonstrate that JNK1 regulates the response to TLR1/2 ligands and suggest a positive feedback loop that may serve to increase the innate immune response to the spirocheteS