Validation of an experimental animal model for corneal additive surgery

Producción CientíficaPurpose: To assess the hen cornea as a model for training and future wound healing studies after implantation of intrastromal corneal ring segments (ICRS) by clinical and optical outcomes. Setting: University of Valladolid, Valladolid, Spain. Design: Experimental study. Methods: One 90°, 150-μm thick polymethyl methacrylate Ferrara ICRS segment was manually implanted at 70-80% depth of 192 Gallus domesticus corneas. Clinical follow-up for 6 months included monitoring corneal thickness, epithelial wound closure, edema, haze, and the location and severity of deposits. The refractive state was also measured. After each animal was euthanized, corneas were processed for direct transmittance and histological analysis. Results: Complications were present in 16% of the eyes. Epithelial wound closure was completed at 3 ± 2 days. A slight corneal edema in the channel site was present for the first 15 days. All corneas had deposits by 4 months located along the inner, outer curvatures and under the segments. Corneal haze was present only at the incision site. ICRS induced hyperopic changes in the refractive state without changes in direct transmitance of central cornea. New cells and extracellular matrix were present around the segment where deposits were seen on clinical follow-up. Conclusions: With hen as an animal model, ICRS were implanted in a precise and reproducible way after a learning curve. Similar to humans, the follow-up period during the first 6 months after implantation showed fast wound closure, deposits, and haze at the incision site. ICRS in hens also reduced the refractive power withoutaffecting the central cornea

Tissue reaction after intrastromal corneal ring implantation in an experimental animal model

Producción CientíficaPurpose To evaluate corneal wound healing in the hen animal model after additive surgery with an intracorneal ring segment (ICRS). Methods We implanted one ICRS in each eye of 76 hens. In control group 1 (n=22 hens), the stromal channel was prepared but no ICRS was inserted. In control group 2 (n=2 hens), no surgery was performed. Animals were randomly separated into groups and euthanized after clinical follow-up of 4 and 12 hours, 1, 2, 3, and 7 days, and 1, 2, 3, 4, and 6 months. Corneas were stained with hematoxylin-eosin. Apoptosis was measured by terminal uridine nick end-labeling assays. Cell proliferation and myofibroblast-like differentiation were assayed by BrdU and α-smooth muscle actin immunofluorescence microscopy. Stromal matrix changes were documented by electron microscopy. Results Epithelial and stromal cell apoptosis around the ICRS-implanted and control group 1 eyes peaked at 12 hours, but continued for 72 hours. In ICRSimplanted eyes, epithelial and stromal proliferation was present at 12 and 24 hours, respectively, and peaked at 7 days and 72 hours, respectively. Some proliferation in the ICRS-implanted group continued through the 6- month follow-up, and myofibroblast-like cells differentiated one to three months after ICRS implantation. The segments rotated within the stroma as the limbal inferior angle approached the epithelium. Conclusions Wound healing after ICRS implantation in hen corneas was similar to that of other corneal surgical wounds in stages. However, there were some specific features related to the small size of the epithelial wound and the device permanently implanted inside the cornea

Resequencing the Vrs1 gene in Spanish barley landraces revealed reversion of six-rowed to two-rowed spike

Six-rowed spike 1 (Vrs1) is a gene of major importance for barley breeding and germplasm management as it is the main gene determining spike row-type (2-rowed vs. 6-rowed). This is a widely used DUS trait, and has been often associated to phenotypic traits beyond spike type. Comprehensive re-sequencing Vrs1 revealed three two-rowed alleles (Vrs1.b2; Vrs1.b3; Vrs1.t1) and four six-rowed (vrs1.a1; vrs1.a2; vrs1.a3; vrs1.a4) in the natural population. However, the current knowledge about Vrs1 alleles and its distribution among Spanish barley subpopulations is still underexploited. We analyzed the gene in a panel of 215 genotypes, made of Spanish landraces and European cultivars. Among 143 six-rowed accessions, 57 had the vrs1.a1 allele, 83 were vrs1.a2, and three showed the vrs1.a3 allele. Vrs1.b3 was found in most two-rowed accessions, and a new allele was observed in 7 out of 50 two-rowed Spanish landraces. This allele, named Vrs1.b5, contains a ‘T’ insertion in exon 2, originally proposed as the causal mutation giving rise to the six-row vrs1.a2 allele, but has an additional upstream deletion that results in the change of 15 amino acids and a potentially functional protein. We conclude that eight Vrs1 alleles (Vrs1.b2, Vrs1.b3, Vrs1.b5, Vrs1.t1, vrs1.a1, vrs1.a2, vrs1.a3, vrs1.a4) discriminate two and six-rowed barleys. The markers described will be useful for DUS identification, plant breeders, and other crop scientists.This work was supported by the Spanish Ministry of Economy, Industry and Competitiveness grants AGL2010-21929, AGL2013-48756-R, RFP2012-00015-00-00, RTA2012-00033-C03-02, and EUI2009-04075 (national code for Plant-KBBE project ExpResBar). CPC was funded by the Spanish Ministry of Economy, Industry and Competitiveness grant no. BES-2011-045905 (linked to project AGL2010-21929). TK and SS were supported by a research fund by the Ministry of Agriculture, Forestry, and Fisheries of Japan (Genomics for Agricultural Innovation grants no. TRS1002). SS was supported by a Grant-in-Aid from the Japan Society for the Promotion of Science (JSPS) Postdoctoral Fellow for Research Abroad and a Grant-in-Aid for Young Scientists (B) (no. 16 K18635)

Candidate genes underlying QTL for flowering time and their interactions in a wide spring barley (Hordeum vulgare L.) cross

Response to vernalization and photoperiod are the main determinants controlling the time to flowering in temperate cereals. While the individual genes that determine a plant's response to these environmental signals are well characterized, the combinatorial effect on flowering time of allelic variants for multiple genes remains unresolved. This study investigated the genetic control of flowering-time in a biparental population of spring barley, derived from a wide cross between a late-flowering European and an early-flowering North-American cultivar. While the major flowering time genes are not segregating in the Beka × Logan cross, large variation in flowering was observed. We identified five QTL, with both parents found to contribute early alleles. The catalog of QTL discovered aligns with several candidate genes affecting flowering time in barley. The combination of particular alleles at HvCEN, HvELF3 and HvFT1 in Logan are responsible for the earliness of this cultivar. Interestingly, earliness for flowering could be further enhanced, with Beka found to contribute three early alleles, including a QTL co-locating with a HvFD-like gene, suggesting that there are diverse aspects of the flowering-time pathway that have been manipulated in these two cultivars. Epistatic interactions between flowering-time QTL or candidate genes were observed in field data and confirmed under controlled conditions. The results of this study link photoperiod-dependent flowering-time genes with earliness per se genes into a single model, thus providing a unique framework that can be used by geneticists and breeders to optimize flowering time in barley.This work was supported by the Spanish Ministry of Economy and Competitiveness (grant numbers AGL2010-21929 and AGL2013-48756-R), the Spanish Ministry of Economy and Competitiveness, the Agencia Estatal de Investigación, and the European Regional Development Fund (grant number AGL2016–80967-R), and Government of Aragon (Research Group A08_20R)

Genetic landscape of 6089 inherited retinal dystrophies affected cases in Spain and their therapeutic and extended epidemiological implications

Inherited retinal diseases (IRDs), defined by dysfunction or progressive loss of photoreceptors, are disorders characterized by elevated heterogeneity, both at the clinical and genetic levels. Our main goal was to address the genetic landscape of IRD in the largest cohort of Spanish patients reported to date. A retrospective hospital-based cross-sectional study was carried out on 6089 IRD affected individuals (from 4403 unrelated families), referred for genetic testing from all the Spanish autonomous communities. Clinical, demographic and familiar data were collected from each patient, including family pedigree, age of appearance of visual symptoms, presence of any systemic findings and geographical origin. Genetic studies were performed to the 3951 families with available DNA using different molecular techniques. Overall, 53.2% (2100/3951) of the studied families were genetically characterized, and 1549 different likely causative variants in 142 genes were identified. The most common phenotype encountered is retinitis pigmentosa (RP) (55.6% of families, 2447/4403). The most recurrently mutated genes were PRPH2, ABCA4 and RS1 in autosomal dominant (AD), autosomal recessive (AR) and X-linked (XL) NON-RP cases, respectively; RHO, USH2A and RPGR in AD, AR and XL for non-syndromic RP; and USH2A and MYO7A in syndromic IRD. Pathogenic variants c.3386G > T (p.Arg1129Leu) in ABCA4 and c.2276G > T (p.Cys759Phe) in USH2A were the most frequent variants identified. Our study provides the general landscape for IRD in Spain, reporting the largest cohort ever presented. Our results have important implications for genetic diagnosis, counselling and new therapeutic strategies to both the Spanish population and other related populations.This work was supported by the Instituto de Salud Carlos III (ISCIII) of the Spanish Ministry of Health (FIS; PI16/00425 and PI19/00321), Centro de Investigación Biomédica en Red Enfermedades Raras (CIBERER, 06/07/0036), IIS-FJD BioBank (PT13/0010/0012), Comunidad de Madrid (CAM, RAREGenomics Project, B2017/BMD-3721), European Regional Development Fund (FEDER), the Organización Nacional de Ciegos Españoles (ONCE), Fundación Ramón Areces, Fundación Conchita Rábago and the University Chair UAM-IIS-FJD of Genomic Medicine. Irene Perea-Romero is supported by a PhD fellowship from the predoctoral Program from ISCIII (FI17/00192). Ionut F. Iancu is supported by a grant from the Comunidad de Madrid (CAM, PEJ-2017-AI/BMD7256). Marta del Pozo-Valero is supported by a PhD grant from the Fundación Conchita Rábago. Berta Almoguera is supported by a Juan Rodes program from ISCIII (JR17/00020). Pablo Minguez is supported by a Miguel Servet program from ISCIII (CP16/00116). Marta Corton is supported by a Miguel Servet program from ISCIII (CPII17/00006). The funders played no role in study design, data collection, data analysis, manuscript preparation and/or publication decisions

Characterisation of two Chinese native Hemicriconemoides species (Nematoda: Criconematidae) with updated descriptions of H. chitwoodi Esser, 1960 and Criconemoides myungsugae Choi & Geraert, 1975

Populations of Hemicriconemoides chitwoodi, H. fujianensis, H. parasinensis, and Criconemoides myungsugae isolated in China from the rhizosphere soil of woody perennials were characterised molecularly, important morphological details being elucidated by SEM observations. The morphometric data of the Chinese populations were compared with all previously reported populations. The SEM observations of en face views indicated that H. chitwoodi and H. parasinensis have lip patterns belonging to type 1 and phylogenetically both species clustered with other Hemicriconemoides species that showed the same kind of lip pattern. On the other hand, H. fujianensis showed a lip pattern belonging to type 2 and phylogenetically appears basal to the above-mentioned clade. SEM observation of C. myungsugae showed that the first lip annulus forms a non-projecting uninterrupted disc and the labial annulus is a rectangular shape with slight dorsal and ventral indentations. Phylogenetic relationships among Criconemoides spp. are apparently not well resolved. The present study provides updated morphological descriptions, molecular diagnostics and phylogenetic relationships of H. chitwoodi, H. fujianensis, H. parasinensis, and C. myungsugae, the last species being the first report from China.Peer reviewe

A 3D Visual Interface for Critiquing-based Recommenders: Architecture and Interaction

Nowadays e-commerce websites offer users such a huge amount of products, which far from facilitating the buying process, actually make it more difficult. Hence, recommenders, which learn from users� preferences, are consolidating as valuable instruments to enhance the buying process in the 2D Web. Indeed, 3D virtual environments are an alternative interface for recommenders. They provide the user with an immersive 3D social experience, enabling a richer visualisation and increasing the interaction possibilities with other users and with the recommender. In this paper, we focus on a novel framework to tightly integrate interactive recommendation systems in a 3D virtual environment. Specifically, we propose to integrate a Collaborative Conversational Recommender (CCR) in a 3D social virtual world. Our CCR Framework defines three layers: the user interaction layer (3D Collaborative Space Client), the communication layer (3D Collaborative Space Server), and the recommendation layer (Collaborative Conversational Recommender). Additionally, we evaluate the framework based on several usability criteria such as learnability, perceived efficiency and effectiveness. Results demonstrate that users positively valued the experience