Bio-Enhanced Neoligaments Graft Bearing FE002 Primary Progenitor Tenocytes: Allogeneic Tissue Engineering & Surgical Proofs-of-Concept for Hand Ligament Regenerative Medicine.

Hand tendon/ligament structural ruptures (tears, lacerations) often require surgical reconstruction and grafting, for the restauration of finger mechanical functions. Clinical-grade human primary progenitor tenocytes (FE002 cryopreserved progenitor cell source) have been previously proposed for diversified therapeutic uses within allogeneic tissue engineering and regenerative medicine applications. The aim of this study was to establish bioengineering and surgical proofs-of-concept for an artificial graft (Neoligaments Infinity-Lock 3 device) bearing cultured and viable FE002 primary progenitor tenocytes. Technical optimization and in vitro validation work showed that the combined preparations could be rapidly obtained (dynamic cell seeding of 10 <sup>5</sup> cells/cm of scaffold, 7 days of co-culture). The studied standardized transplants presented homogeneous cellular colonization in vitro (cellular alignment/coating along the scaffold fibers) and other critical functional attributes (tendon extracellular matrix component such as collagen I and aggrecan synthesis/deposition along the scaffold fibers). Notably, major safety- and functionality-related parameters/attributes of the FE002 cells/finished combination products were compiled and set forth (telomerase activity, adhesion and biological coating potentials). A two-part human cadaveric study enabled to establish clinical protocols for hand ligament cell-assisted surgery (ligamento-suspension plasty after trapeziectomy, thumb metacarpo-phalangeal ulnar collateral ligamentoplasty). Importantly, the aggregated experimental results clearly confirmed that functional and clinically usable allogeneic cell-scaffold combination products could be rapidly and robustly prepared for bio-enhanced hand ligament reconstruction. Major advantages of the considered bioengineered graft were discussed in light of existing clinical protocols based on autologous tenocyte transplantation. Overall, this study established proofs-of-concept for the translational development of a functional tissue engineering protocol in allogeneic musculoskeletal regenerative medicine, in view of a pilot clinical trial

Diversity arrays technology (DArT) markers in apple for genetic linkage maps

Diversity Arrays Technology (DArT) provides a high-throughput whole-genome genotyping platform for the detection and scoring of hundreds of polymorphic loci without any need for prior sequence information. The work presented here details the development and performance of a DArT genotyping array for apple. This is the first paper on DArT in horticultural trees. Genetic mapping of DArT markers in two mapping populations and their integration with other marker types showed that DArT is a powerful high-throughput method for obtaining accurate and reproducible marker data, despite the low cost per data point. This method appears to be suitable for aligning the genetic maps of different segregating populations. The standard complexity reduction method, based on the methylation-sensitive PstI restriction enzyme, resulted in a high frequency of markers, although there was 52–54% redundancy due to the repeated sampling of highly similar sequences. Sequencing of the marker clones showed that they are significantly enriched for low-copy, genic regions. The genome coverage using the standard method was 55–76%. For improved genome coverage, an alternative complexity reduction method was examined, which resulted in less redundancy and additional segregating markers. The DArT markers proved to be of high quality and were very suitable for genetic mapping at low cost for the apple, providing moderate genome coverage

Eficiência de fungicidas para o controle da ferrugem-asiática da soja, Phakopsora pachyrhizi, na safra 2011/12: resultados sumarizados dos ensaios cooperativos.

Material e método: Tabela 1 - Instituições, locais, cultivares, datas da semeadura da soja, presença (1) ou ausência (0) de sintomas de ferrugem na primeira aplicação de fungicida (incidência - INC). Tabela 2 - Ingrediente ativo (i.a.), produto comercial (p.c.) e dose dos fungicidas nos tratamentos para o controle da ferrugem-asiática da soja, safra 2011/12. Resultados e discussões: Figura 1 - Box plot da porcentagem de controle da ferrugem, para os diferentes tratamentos com fungicidas, em relação à média da severidade da testemunha sem controle (T1). Figura 2 - Box plot da porcentagem de redução da produtividade, para os diferentes tratamentos com fungicidas, em relação à média da produtividade do melhor tratamento. Tabela 3: Severidade da ferruge, porcentagem de controle em relação à testemunha sem tratamento, produtividade (Kg ha-1) e porcentagem de redução de produtividade (RP) em relação ao tratamento com a maior produtividade, para os diferentes tratamentos. Média de 11 ensaios para severidade e de 15 ensaios para produtividade. Rede de ensaios cooperativos, safra 2011/12.bitstream/item/61841/1/CT93.2012.pd

Linkage map construction involving a reciprocal translocation

This paper is concerned with a novel statistical–genetic approach for the construction of linkage maps in populations obtained from reciprocal translocation heterozygotes of barley (Hordeum vulgare L.). Using standard linkage analysis, translocations usually lead to ‘pseudo-linkage’: the mixing up of markers from the chromosomes involved in the translocation into a single linkage group. Close to the translocation breakpoints recombination is severely suppressed and, as a consequence, ordering markers in those regions is not feasible. The novel strategy presented in this paper is based on (1) disentangling the “pseudo-linkage” using principal coordinate analysis, (2) separating individuals into translocated types and normal types and (3) separating markers into those close to and those more distant from the translocation breakpoints. The methods make use of a consensus map of the species involved. The final product consists of integrated linkage maps of the distal parts of the chromosomes involved in the translocation

Deflection control for reinforced recycled aggregate concrete beams: Experimental database and extension of the fib Model Code 2010 model

Recycled aggregate concrete (RAC) has emerged as a viable solution for solving some of the environmental problems of concrete production. However, design guidelines for deflection control of reinforced RAC members have not yet been proposed. This study presents a comprehensive analysis of the applicability of the fib Model Code 2010 (MC2010) deflection control model to reinforced RAC beams. Three databases of long-term studies on natural aggregate concrete (NAC) and RAC beams were compiled and meta-analyses of deflection predictions by MC2010 were performed. First, the MC2010 deflection control model was tested against a large database of long-term tests on NAC beams. Second, a database of RAC and companion NAC beams was compiled and initial and long-term deflections were calculated using the MC2010 model. It was shown that deflections of RAC beams are significantly underestimated relative to NAC beams. Previously proposed modifications for MC2010 equations for shrinkage strain and creep coefficient were used, and new modifications for the modulus of elasticity and empirical coefficient β were proposed. The improved MC2010 deflection control model on RAC beams was shown to have equal performance to that on companion NAC beams. The proposals presented in this paper can help engineers to more reliably perform deflection control of reinforced RAC members.This is the peer-reviewed version of the article: N. Tošić, S. Marinković, and J. de Brito, ‘Deflection control for reinforced recycled aggregate concrete beams: Experimental database and extension of the fib Model Code 2010 model’, Structural Concrete, vol. 20, no. 6, pp. 2015–2029, 2019 [https://doi.org/10.1002/suco.201900035

Association mapping of spot blotch resistance in wild barley

Spot blotch, caused by Cochliobolus sativus, is an important foliar disease of barley. The disease has been controlled for over 40 years through the deployment of cultivars with durable resistance derived from the line NDB112. Pathotypes of C. sativus with virulence for the NDB112 resistance have been detected in Canada; thus, many commercial cultivars are vulnerable to spot blotch epidemics. To increase the diversity of spot blotch resistance in cultivated barley, we evaluated 318 diverse wild barley accessions comprising the Wild Barley Diversity Collection (WBDC) for reaction to C. sativus at the seedling stage and utilized an association mapping (AM) approach to identify and map resistance loci. A high frequency of resistance was found in the WBDC as 95% (302/318) of the accessions exhibited low infection responses. The WBDC was genotyped with 558 Diversity Array Technology (DArT®) and 2,878 single nucleotide polymorphism (SNP) markers and subjected to structure analysis before running the AM procedure. Thirteen QTL for spot blotch resistance were identified with DArT and SNP markers. These QTL were found on chromosomes 1H, 2H, 3H, 5H, and 7H and explained from 2.3 to 3.9% of the phenotypic variance. Nearly half of the identified QTL mapped to chromosome bins where spot blotch resistance loci were previously reported, offering some validation for the AM approach. The other QTL mapped to unique genomic regions and may represent new spot blotch resistance loci. This study demonstrates that AM is an effective technique for identifying and mapping QTL for disease resistance in a wild crop progenitor

Diversity Arrays Technology (DArT) for Pan-Genomic Evolutionary Studies of Non-Model Organisms

Background: High-throughput tools for pan-genomic study, especially the DNA microarray platform, have sparked a remarkable increase in data production and enabled a shift in the scale at which biological investigation is possible. The use of microarrays to examine evolutionary relationships and processes, however, is predominantly restricted to model or near-model organisms. Methodology/Principal Findings: This study explores the utility of Diversity Arrays Technology (DArT) in evolutionary studies of non-model organisms. DArT is a hybridization-based genotyping method that uses microarray technology to identify and type DNA polymorphism. Theoretically applicable to any organism (even one for which no prior genetic data are available), DArT has not yet been explored in exclusively wild sample sets, nor extensively examined in a phylogenetic framework. DArT recovered 1349 markers of largely low copy-number loci in two lineages of seed-free land plants: the diploid fern Asplenium viride and the haploid moss Garovaglia elegans. Direct sequencing of 148 of these DArT markers identified 30 putative loci including four routinely sequenced for evolutionary studies in plants. Phylogenetic analyses of DArT genotypes reveal phylogeographic and substrate specificity patterns in A. viride, a lack of phylogeographic pattern in Australian G. elegans, and additive variation in hybrid or mixed samples. Conclusions/Significance: These results enable methodological recommendations including procedures for detecting and analysing DArT markers tailored specifically to evolutionary investigations and practical factors informing the decision to use DArT, and raise evolutionary hypotheses concerning substrate specificity and biogeographic patterns. Thus DArT is a demonstrably valuable addition to the set of existing molecular approaches used to infer biological phenomena such as adaptive radiations, population dynamics, hybridization, introgression, ecological differentiation and phylogeography

Eficiência de fungicidas para o controle da ferrugem-asiática da soja, Phakopsora pachyrhizi, na safra 2010/11: resultados sumarizados dos ensaios cooperativos.

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