Participatory plant breeding: a way to arrive at better-adapted onion varieties

The search for varieties that are better adapted to organic farming is a current topic in the organic sector. Breeding programmes specific for organic agriculture should solve this problem. Collaborating with organic farmers in such programmes, particularly in the selection process, can potentially result in varieties better adapted to their needs. Here, we assume that organic farmers' perceptive of plant health is broader than that of conventional breeders. Two organic onion farmers and one conventional onion breeder were monitored in their selection activities in 2004 and 2005 in order to verify whether and in which way this broader view on plant health contributes to improvement of organic varieties. They made selections by positive mass selection in three segregating populations under organic conditions. The monitoring showed that the organic farmers selected in the field for earliness and downy mildew and after storage for bulb characteristics. The conventional breeder selected only after storage. Farmers and breeder applied identical selection directions for bulb traits as a round shape, better hardness and skin firmness. This resulted in smaller bulbs in the breeders’ populations, while the bulbs in the farmer populations were bigger than in the original population. In 2006 and 2007 the new onion populations will be compared with each other and the original populations to determine the selection response

Caby Photometry of the Hyades: Comparisons to the Field Stars

Intermediate-band photometry of the Hyades cluster on the Caby system is presented for dwarf stars ranging from spectral type A through late K. A mean hk, b-y relation is constructed using only single stars without anomalous atmospheres and compared to the field stars of the solar neighborhood. For the F dwarfs, the Hyades relation defines an approximate LOWER bound in the two-color diagram, consistent with an [Fe/H] between +0.10 and +0.15. These index-color diagrams follow the common convention of presenting stars with highest abundance at the bottom of the plot although the index values for the metal-rich stars are numerically larger. For field F dwarfs in the range [Fe/H] between +0.4 and -1.0, [Fe/H] = -5.6 delta-hk + 0.125, with no evidence for a color dependence in the slope. For the G and K dwarfs, the Hyades mean relation crosses the field star distribution in the two-color diagram, defining an approximate UPPER bound for the local disk stars. Stars found above the Hyades stars fall in at least one of three categories: [Fe/H] below -0.7, [Fe/H] above that of the Hyades, or chromospherically active. It is concluded that, contrary to the predictions of model atmospheres, the hk index for cool dwarfs at a given color hits a maximum value for stars below solar composition and, with increasing [Fe/H] above some critical value, declines. This trend is consistent, however, with the predictions from synthetic indices based upon much narrower Ca filters where the crossover is caused by the metallicity sensitivity of b-y.Comment: 13 pages, 9 eps figures, 1 tex table, 1 ascii tabl

Gene editing of PKLR gene in human hematopoietic progenitors through 5' and 3' UTR modified TALEN mRNA

TheauthorswouldliketothankMiguelA.MartinforthecarefulmaintenanceofNSGmice,andRebecaSa ́nchezandOmairaAlberquillafortheirtechnicalassistanceinflowcytometry.TheauthorsalsothankFundacio ́n Botı ́n forpromotingtranslationalresearchattheHemato-poieticInnovativeTherapiesDivisionoftheCIEMATPyruvate Kinase Deficiency (PKD) is a rare erythroid metabolic disease caused by mutations in the PKLR gene, which encodes the erythroid specific Pyruvate Kinase enzyme. Erythrocytes from PKD patients show an energetic imbalance and are susceptible to hemolysis. Gene editing of hematopoietic stem cells (HSCs) would provide a therapeutic benefit and improve safety of gene therapy approaches to treat PKD patients. In previous studies, we established a gene editing protocol that corrected the PKD phenotype of PKD-iPSC lines through a TALEN mediated homologous recombination strategy. With the goal of moving toward more clinically relevant stem cells, we aim at editing the PKLR gene in primary human hematopoietic progenitors and hematopoietic stem cells (HPSCs). After nucleofection of the gene editing tools and selection with puromycin, up to 96% colony forming units showed precise integration. However, a low yield of gene edited HPSCs was associated to the procedure. To reduce toxicity while increasing efficacy, we worked on i) optimizing gene editing tools and ii) defining optimal expansion and selection times. Different versions of specific nucleases (TALEN and CRISPR-Cas9) were compared. TALEN mRNAs with 5' and 3' added motifs to increase RNA stability were the most efficient nucleases to obtain high gene editing frequency and low toxicity. Shortening ex vivo manipulation did not reduce the efficiency of homologous recombination and preserved the hematopoietic progenitor potential of the nucleofected HPSCs. Lastly, a very low level of gene edited HPSCs were detected after engraftment in immunodeficient (NSG) mice. Overall, we showed that gene editing of the PKLR gene in HPSCs is feasible, although further improvements must to be done before the clinical use of the gene editing to correct PKD.S

Manipulation and removal of defects in spontaneous optical patterns

Defects play an important role in a number of fields dealing with ordered structures. They are often described in terms of their topology, mutual interaction and their statistical characteristics. We demonstrate theoretically and experimentally the possibility of an active manipulation and removal of defects. We focus on the spontaneous formation of two-dimensional spatial structures in a nonlinear optical system, a liquid crystal light valve under single optical feedback. With increasing distance from threshold, the spontaneously formed hexagonal pattern becomes disordered and contains several defects. A scheme based on Fourier filtering allows us to remove defects and to restore spatial order. Starting without control, the controlled area is progressively expanded, such that defects are swept out of the active area.Comment: 4 pages, 4 figure

Upregulation of NKG2D ligands impairs hematopoietic stem cell function in Fanconi anemia

Altres ajuts: Fondo Europeo de Desarrollo Regional (FEDER); Next Generation EU; EUROFANCOLEN); Comunidad de Madrid (AvanCell, B2017/BMD-3692); ICREA-Academia program.Fanconi anemia (FA) is the most prevalent inherited bone marrow failure (BMF) syndrome. Nevertheless, the pathophysiological mechanisms of BMF in FA have not been fully elucidated. Since FA cells are defective in DNA repair, we hypothesized that FA hematopoietic stem and progenitor cells (HSPCs) might express DNA damage-associated stress molecules such as natural killer group 2 member D ligands (NKG2D-Ls). These ligands could then interact with the activating NKG2D receptor expressed in cytotoxic NK or CD8+ T cells, which may result in progressive HSPC depletion. Our results indeed demonstrated upregulated levels of NKG2D-Ls in cultured FA fibroblasts and T cells, and these levels were further exacerbated by mitomycin C or formaldehyde. Notably, a high proportion of BM CD34+ HSPCs from patients with FA also expressed increased levels of NKG2D-Ls, which correlated inversely with the percentage of CD34+ cells in BM. Remarkably, the reduced clonogenic potential characteristic of FA HSPCs was improved by blocking NKG2D-NKG2D-L interactions. Moreover, the in vivo blockage of these interactions in a BMF FA mouse model ameliorated the anemia in these animals. Our study demonstrates the involvement of NKG2D-NKG2D-L interactions in FA HSPC functionality, suggesting an unexpected role of the immune system in the progressive BMF that is characteristic of FA

International Children’s Rights: Reflections on a Complex, Dynamic, and Relatively Young Area of Law

This chapter reflects on the aim of the International Children’s Rights volume to provide those wishing to study, research, and practice international children’s rights law with a contemporary and comprehensive legal text. It recaps on the themes that emerged from the process of commissioning and editing the various contributions from some of the world’s leading and emerging legal scholars in the area of children’s rights. It marks the progress that has been made in the implementation of children’s rights law and the many challenges that still exist in the implementation of the CRC and associated international instruments. It notes that legal scholarship in the field of children’s rights is still developing and that, although multidisciplinary research and analysis is valuable, it is important to reaffirm children’s rights as a field of law and legal practice. International children’s rights is a complex, dynamic, and relatively young area of law. As the contributions to the collection show, it is diverse and evolving, with many new aspects and issues worthy of analysis and scrutiny. This chapter encapsulates the aspiration of the volume editors that the book contribute to the scrutiny of the legal implications of the CRC, recognizing the unique features of international children’s rights law, adding to the ongoing development of this important area of law.Effective Protection of Fundamental Rights in a pluralist worl

The C-Terminal Domain of the Arabinosyltransferase Mycobacterium tuberculosis EmbC Is a Lectin-Like Carbohydrate Binding Module

The D-arabinan-containing polymers arabinogalactan (AG) and lipoarabinomannan (LAM) are essential components of the unique cell envelope of the pathogen Mycobacterium tuberculosis. Biosynthesis of AG and LAM involves a series of membrane-embedded arabinofuranosyl (Araf) transferases whose structures are largely uncharacterised, despite the fact that several of them are pharmacological targets of ethambutol, a frontline drug in tuberculosis therapy. Herein, we present the crystal structure of the C-terminal hydrophilic domain of the ethambutol-sensitive Araf transferase M. tuberculosis EmbC, which is essential for LAM synthesis. The structure of the C-terminal domain of EmbC (EmbCCT) encompasses two sub-domains of different folds, of which subdomain II shows distinct similarity to lectin-like carbohydrate-binding modules (CBM). Co-crystallisation with a cell wall-derived di-arabinoside acceptor analogue and structural comparison with ligand-bound CBMs suggest that EmbCCT contains two separate carbohydrate binding sites, associated with subdomains I and II, respectively. Single-residue substitution of conserved tryptophan residues (Trp868, Trp985) at these respective sites inhibited EmbC-catalysed extension of LAM. The same substitutions differentially abrogated binding of di- and penta-arabinofuranoside acceptor analogues to EmbCCT, linking the loss of activity to compromised acceptor substrate binding, indicating the presence of two separate carbohydrate binding sites, and demonstrating that subdomain II indeed functions as a carbohydrate-binding module. This work provides the first step towards unravelling the structure and function of a GT-C-type glycosyltransferase that is essential in M. tuberculosis. Author Summary Top Tuberculosis (TB), an infectious disease caused by the bacillus Mycobacterium tuberculosis, burdens large swaths of the world population. Treatment of active TB typically requires administration of an antibiotic cocktail over several months that includes the drug ethambutol. This front line compound inhibits a set of arabinosyltransferase enzymes, called EmbA, EmbB and EmbC, which are critical for the synthesis of arabinan, a vital polysaccharide in the pathogen's unique cell envelope. How precisely ethambutol inhibits arabinosyltransferase activity is not clear, in part because structural information of its pharmacological targets has been elusive. Here, we report the high-resolution structure of the C-terminal domain of the ethambutol-target EmbC, a 390-amino acid fragment responsible for acceptor substrate recognition. Combining the X-ray crystallographic analysis with structural comparisons, site-directed mutagenesis, activity and ligand binding assays, we identified two regions in the C-terminal domain of EmbC that are capable of binding acceptor substrate mimics and are critical for activity of the full-length enzyme. Our results begin to define structure-function relationships in a family of structurally uncharacterised membrane-embedded glycosyltransferases, which are an important target for tuberculosis therapy