Fibroblasts activation and abnormal extracellular matrix remodelling as common hallmarks in three cancer-prone genodermatoses

Background. Recessive dystrophic epidermolysis bullosa (RDEB), Kindler syndrome (KS) and xeroderma pigmentosum complementation group C (XPC) are three cancer-prone genodermatoses whose causal genetic mutations cannot fully explain, on their own, the array of associated phenotypic manifestations. Recent evidence highlights the role of the stromal microenvironment in the pathology of these disorders. Objectives. To investigate, by means of comparative gene expression analysis, the role played by dermal fibroblasts in the pathogenesis of RDEB, KS and XPC. Methods. We conducted RNA-Seq analysis, which included a thorough examination of the differentially expressed genes, a functional enrichment analysis and a description of affected signalling circuits. Transcriptomic data were validated at the protein level in cell cultures, serum samples and skin biopsies. Results. Interdisease comparisons against control fibroblasts revealed a unifying signature of 186 differentially expressed genes and four signalling pathways in the three genodermatoses. Remarkably, some of the uncovered expression changes suggest a synthetic fibroblast phenotype characterized by the aberrant expression of extracellular matrix (ECM) proteins. Western blot and immunofluorescence in situ analyses validated the RNA-Seq data. In addition, enzyme-linked immunosorbent assay revealed increased circulating levels of periostin in patients with RDEB. Conclusions. Our results suggest that the different causal genetic defects converge into common changes in gene expression, possibly due to injury-sensitive events. These, in turn, trigger a cascade of reactions involving abnormal ECM deposition and underexpression of antioxidant enzymes. The elucidated expression signature provides new potential biomarkers and common therapeutic targets in RDEB, XPC and KS.This study was supported by grants from the Spanish Ministry of Economy and Competitiveness (SAF2013-43475R, SAF2017-88908-R and SAF2017-86810-R); from Instituto de Salud Carlos III and CIBERER, cofunded with European Regional Development Funds (ERDF) (PT13/0001/0007, PI14/00931, PI15/00716, PI15/00956, PT17/0009/0006 and PI17/01747); and from the European Union (HEALTH-F2-2011-261392 and H2020-INFRADEV-1-2015-1/ELIXIR-EXCELERATEref. 676559). Additional funding from Comunidad de Madrid (AvanCell-CM S2017/BMD-3692); Catalan Government (AGAUR 2014_SGR_603); ‘Fundacio' La Marató de TV3, 01331-30’; CERCA Programme/Generalitat de Catalunya; and ‘Fundación Científica de la Asociación Española Contra el Cáncer’, Spain

Evaluation of growth in diesel fuel and surfactants production ability by bacteria isolated from fuels in Costa Rica

Abstract: A total of 149 bacterial strains previously isolated from fuels stored in Costa Rica were selected in terms of their ability to grow aerobically in diesel and produce bioemulsifier active compounds. The diesel growth was evaluated by a redox-indicator based test, and surfactant production was estimated indirectly by both the emulsification index determination (E24) and hemolytic activity. Twenty-six strains (16.8%) were considered as capable of growing in diesel, while surfactant production was detected in 22 (14.8%), estimated according to E24. Seven strains showed high production of biosurfactants (E24 ≥ 50%), headed by Pseudomonas aeruginosa 148D-O, P. aeruginosa 87R-B and Bacillus pumilus 133S-B. No significant correlation was observed between hemolytic patterns and growth outcomes in diesel or E24. Surfactant producing strains should be studied further to assess its potential applications.Evaluación de crecimiento en combustible diesel y capacidad de producción de surfactantes en bacterias aisladas de combustibles en Costa Rica Resumen: Un total de 149 cepas bacterianas previamente aisladas de combustibles almacenados en Costa Rica fueron seleccionadas en términos de sus habilidades para crecer aeróbicamente en diesel y producir compuestos con actividad bioemulsificante. El crecimiento en diesel fue evaluado por medio de un test basado en un indicador redox, y la producción de surfactantes fue estimada indirectamente con las determinaciones del índice de emulsificación (E24) y la actividad hemolítica. Veintiseis cepas (16,8%) fueron consideradas como capaces de crecer en diesel, mientras que la producción de surfactantes fue detectada en 22 (14,8%), estimado de acuerdo con el E24. Siete cepas mostraron alta producción de biosurfactantes (E24 ≥ 50%), encabezadas por Pseudomonas aeruginosa 148D-O, P. aeruginosa 87R-B y Bacillus pumilus 133S-B. No se observó correlación significativa entre los patrones de hemólisis y los resultados de crecimiento en diesel o E24. Las cepas productoras de surfactantes deben ser estudiadas más a fondo para evaluar sus potenciales aplicaciones

A first checklist to the vascular plants of La Amistad International Park (PILA), Costa Rica-Panama.

La Amistad International Park is a World Heritage Site, which comprises 401,000 ha of mainly upland continuous natural vegetation straddling the Costa Rica and Panama border. We present a first checklist of vascular plant diversity for the park and a brief discussion of how this diversity is distributed by elevation and vegetation type together with a superficial assessment of floristic affinities. The checklist recognises 3,046 vascular plant species, 26 of which are lycopods, 433 are ferns and 2,586 are seed plants. Of these, 16 are new records for Costa Rica and 39 are for the flora of Panama; 14 represent undescribed or new species to science and 73 are endemic to La Amistad or its buffer zone. For each species we document its presence within the Park by citing herbarium specimens and the associated elevational range, together with their global distribution, extinction risk assessments where undertaken, whether the taxon is exotic

Efficient CRISPR-Cas9-Mediated Gene Ablation in Human Keratinocytes to Recapitulate Genodermatoses: Modeling of Netherton Syndrome

Current efforts to find specific genodermatoses treatments and define precise pathogenesis mechanisms require appropriate surrogate models with human cells. Although transgenic and gene knockout mouse models for several of these disorders exist, they often fail to faithfully replicate the clinical and histopathological features of the human skin condition. We have established a highly efficient method for precise deletion of critical gene sequences in primary human keratinocytes, based on CRISPR-Cas9-mediated gene editing. Using this methodology, in the present study we generated a model of Netherton syndrome by disruption of SPINK5. Gene-edited cells showed absence of LEKTI expression and were able to recapitulate a hyperkeratotic phenotype with most of the molecular hallmarks of Netherton syndrome, after grafting to immunodeficient mice and in organotypic cultures. To validate the model as a platform for therapeutic intervention, we tested an ex vivo gene therapy approach using a lentiviral vector expressing SPINK5. Re-expression of SPINK5 in an immortalized clone of SPINK5-knockout keratinocytes was capable of reverting from Netherton syndrome to a normal skin phenotype in vivo and in vitro. Our results demonstrate the feasibility of modeling genodermatoses, such as Netherton syndrome, by efficiently disrupting the causative gene to better understand its pathogenesis and to develop novel therapeutic approaches