Systematic Collaborative Reanalysis of Genomic Data Improves Diagnostic Yield in Neurologic Rare Diseases

Altres ajuts: Generalitat de Catalunya, Departament de Salut; Generalitat de Catalunya, Departament d'Empresa i Coneixement i CERCA Program; Ministerio de Ciencia e Innovación; Instituto Nacional de Bioinformática; ELIXIR Implementation Studies (CNAG-CRG); Centro de Investigaciones Biomédicas en Red de Enfermedades Raras; Centro de Excelencia Severo Ochoa; European Regional Development Fund (FEDER).Many patients experiencing a rare disease remain undiagnosed even after genomic testing. Reanalysis of existing genomic data has shown to increase diagnostic yield, although there are few systematic and comprehensive reanalysis efforts that enable collaborative interpretation and future reinterpretation. The Undiagnosed Rare Disease Program of Catalonia project collated previously inconclusive good quality genomic data (panels, exomes, and genomes) and standardized phenotypic profiles from 323 families (543 individuals) with a neurologic rare disease. The data were reanalyzed systematically to identify relatedness, runs of homozygosity, consanguinity, single-nucleotide variants, insertions and deletions, and copy number variants. Data were shared and collaboratively interpreted within the consortium through a customized Genome-Phenome Analysis Platform, which also enables future data reinterpretation. Reanalysis of existing genomic data provided a diagnosis for 20.7% of the patients, including 1.8% diagnosed after the generation of additional genomic data to identify a second pathogenic heterozygous variant. Diagnostic rate was significantly higher for family-based exome/genome reanalysis compared with singleton panels. Most new diagnoses were attributable to recent gene-disease associations (50.8%), additional or improved bioinformatic analysis (19.7%), and standardized phenotyping data integrated within the Undiagnosed Rare Disease Program of Catalonia Genome-Phenome Analysis Platform functionalities (18%)

Expanding the recombinant protein quality in Lactococcus lactis

Background: Escherichia coli has been a main host for the production of recombinant proteins of biomedical interest, but conformational stress responses impose severe bottlenecks that impair the production of soluble, proteolytically stable versions of many protein species. In this context, emerging Generally Recognized As Safe (GRAS) bacterial hosts provide alternatives as cell factories for recombinant protein production, in which limitations associated to the use of Gram-negative microorganisms might result minimized. Among them, Lactic Acid Bacteria and specially Lactococcus lactis are Gram-positive GRAS organisms in which recombinant protein solubility is generically higher and downstream facilitated, when compared to E. coli. However, deep analyses of recombinant protein quality in this system are still required to completely evaluate its performance and potential for improvement. - Results : we have explored here the conformational quality (through specific fluorescence emission) and solubility of an aggregation-prone GFP variant (VP1GFP) produced in L. lactis. In this context, our results show that parameters such as production time, culture conditions and growth temperature have a dramatic impact not only on protein yield, but also on protein solubility and conformational quality, that are particularly favored under fermentative metabolism. - Conclusions: metabolic regime and cultivation temperature greatly influence solubility and conformational quality of an aggregation-prone protein in L. lactis. Specifically, the present study proves that anaerobic growth is the optimal condition for recombinant protein production purposes. Besides, growth temperature plays an important role regulating both protein solubility and conformational quality. Additionally, our results also prove the great versatility for the manipulation of this bacterial system regarding the improvement of functionality, yield and quality of recombinant proteins in this species. These findings not only confirm L. lactis as an excellent producer of recombinant proteins but also reveal room for significant improvement by the exploitation of external protein quality modulators

Fluorescent dye labeling changes the biodistribution of cell-targeted nanoparticles

Fluorescent dye labeling is a common strategy to analyze the fate of administered nanoparticles in living organisms. However, to which extent the labeling processes can alter the original nanoparticle biodistribution has been so far neglected. In this work, two widely used fluorescent dye molecules, namely, ATTO488 (ATTO) and Sulfo-Cy5 (S-Cy5), have been covalently attached to a well-characterized CXCR4-targeted self-assembling protein nanoparticle (known as T22-GFP-H6). The biodistribution of labeled T22-GFP-H6-ATTO and T22-GFP-H6-S-Cy5 nanoparticles has been then compared to that of the non-labeled nanoparticle in different CXCR4+ tumor mouse models. We observed that while parental T22-GFP-H6 nanoparticles accumulated mostly and specifically in CXCR4+ tumor cells, labeled T22-GFP-H6-ATTO and T22-GFP-H6-S-Cy5 nanoparticles showed a dramatic change in the biodistribution pattern, accumulating in non-target organs such as liver or kidney while reducing tumor targeting capacity. Therefore, the use of such labeling molecules should be avoided in target and non-target tissue uptake studies during the design and development of targeted nanoscale drug delivery systems, since their effect over the fate of the nanomaterial can lead to considerable miss-interpretations of the actual nanoparticle biodistribution

Guia de pràctica clínica sobre el maneig i tractament d’úlceres d’extremitats inferiors

Úlceres; Extremitats inferiors; Cures; EpidemiologiaÚlceras; Extremidades inferiores; Curas; EpidemiologíaUlcers; Lower extremities; Cures; EpidemiologyLes úlceres cròniques en les extremitats inferiors són un problema de salut amb importants repercussions socioeconòmiques a causa de la seva llarga evolució. Pot minvar la qualitat de vida del pacient i propiciar l’absentisme laboral, i constitueix un gran repte per als professionals sanitaris. L'objectiu d'aquest document és oferir al professional un coneixement actualitzat sobre les millors actuacions preventives i curatives en el maneig i tractament de les úlceres d’extremitat inferior. També facilitar informació sobre mesures diagnòstiques i terapèutiques per a cada situació clínica, per poder millorar la qualitat i eficiència de les cures proporcionades des d'una perspectiva holística i individualitzada

Guia de pràctica clínica sobre el maneig i tractament d’úlceres d’extremitats inferiors

Úlceres; Extremitats inferiors; Cures; EpidemiologiaÚlceras; Extremidades inferiores; Curas; EpidemiologíaUlcers; Lower extremities; Cures; EpidemiologyLes úlceres cròniques en les extremitats inferiors són un problema de salut amb importants repercussions socioeconòmiques a causa de la seva llarga evolució. Pot minvar la qualitat de vida del pacient i propiciar l’absentisme laboral, i constitueix un gran repte per als professionals sanitaris. L'objectiu d'aquest document és oferir al professional un coneixement actualitzat sobre les millors actuacions preventives i curatives en el maneig i tractament de les úlceres d’extremitat inferior. També facilitar informació sobre mesures diagnòstiques i terapèutiques per a cada situació clínica, per poder millorar la qualitat i eficiència de les cures proporcionades des d'una perspectiva holística i individualitzada