Filaggrin gene mutations and new SNPs in asthmatic patients: a cross-sectional study in a Spanish population

[EN] Background: Several null-mutations in the FLG gene that produce a decrease or absence of filaggrin in the skin and predispose to atopic dermatitis and ichthyosis vulgaris have been described. The relationship with asthma is less clear and may be due to the influence of atopy in patients with associated asthma. Methods: Four hundred individuals were included, 300 patients diagnosed with asthma divided into two groups according to their phenotype (allergic and non-allergic asthma) and 100 strictly characterized controls. The coding region and flanking regions of the FLG gene were amplified by PCR. We proceeded to the characterization of potential gene variants in that region by RFLP and sequencing and analysed their association with lung function parameters, asthma control and severity, and quality of life. Results: We identified two null-mutations (R501X and 2282del4), seven SNPs previously described in databases and three SNPs that had not been previously described. One of the SNP identified in this study (1741A > T) was more frequently detected in patients with non-allergic asthma, worse FVC, FEV1 and PEF values and a higher treatment step. In addition, lowered spirometric values were observed in the non-allergic group carrying any of the nonsynonymous SNPs. Conclusions: In the association study of genetic variants of the FLG gene in our population the 1741A > T polymorphism seems to be associated with non-allergic asthma

Análisis y caracterización de genes de Botrytis cinerea cuya expresión se induce in planta en la interacción B.cinerea-tomate

[ES] Botrytis cinerea es un hongo fitopatógeno causante de la podredumbre gris y considerado como uno de los principales microosganismos responsables del deterioro de frutas y hortalizas durante su cultivo y en postcosecha y, por tanto, responsable de grandes pérdidas económicas. Es un patógeno generalista que puede atacar especies de la mayoría de las familias de dicotiledóneas. B. cinerea es un hongo necrotrofo que requiere la muerte de las células vegetales para poder alimentarse y desarrollar la infección. Sin embargo, los mecanismos que participan en estos procesos aún no han sido esclarecidos en su totalidad. Con el objeto de contribuir a la caracterización de los mecanismos de patogenicidad de B.cinerea, se procedió a llevar a cabo un análisis de expresión génica diferencial durante la interacción B.cinerea-tomate para tratar de identificar genes cuya expresión se inducía, o aumentaba significativamente, durante la interacción del patógeno con su huésped. Esta aproximación se basa en la consideración de que la inducción de la expresión de un gen en unas condiciones dadas o en un proceso particular nos permite presuponer que dicho gen y su producto génico juegan un papel importante en el proceso analizado o son necesarios para la adaptación del organismo a las condiciones concretas estudiadas. Fruto de la aproximación experimental aplicada se aislaron varios fragmentos de cDNA derivados de genes cuya expresión se inducía, o aumentaba significativamente, in planta, lo que hace suponer que los genes correspondientes están implicados en el proceso de infección. Esta tesis aborda la caracterización de dos de esos fragmentos de cDNA.[EN] Botrytis cinerea is a phytopathogenic fungus that causes gray mold and considered one of the main microosganismos responsible for the deterioration of fruits and vegetables during cultivation and post harvest and, therefore, responsible for great economic losses. It is a generalist pathogen species can attack most of the families of flowering plants. B. cinerea is a necrotrophic fungus that requires the death of plant cells to feed and develop the infection. However, the mechanisms involved in these processes have not yet been fully clarified. In order to contribute to the characterization of the mechanisms of pathogenicity of B.cinerea, proceeded to carry out an analysis of differential gene expression during the interaction B.cinerea-tomato to try to identify genes whose expression was induced, or increased significantly during the interaction of the pathogen with its host. This approach is based on the consideration that the induction of gene expression under given conditions or in a particular process allows us to assume that such gene and its gene product plays an important role in the process under consideration or are necessary for adaptation the organism to the specific conditions studied. Fruit of the experimental approach applied were isolated cDNA fragments from genes whose expression was induced or increased significantly, in planta, which suggests that the corresponding genes are involved in the infection process. This thesis addresses the characterization of two of these cDNA fragments

Anterior cruciate ligament innervation in primary knee osteoarthritis

Objective. To relate the Anterior Cruciate Ligament (ACL) innervation and histologic degeneration status to the knee osteoarthritis radiologic and functional status. Design. Prospective observational study including 30 consecutive patients affected by primary knee osteoarthritis undergoing Total Knee Arthroplasty (TKA). All patients suffering secondary knee osteoarthritis, an antecedent of an infectious process, malignant process, autoimmune disorder, or previous knee surgery were excluded. We recorded biodemographic, clinical, and radiologic variables of all participants previous to the TKA procedure. ACL tissue was harvested during TKA standard procedure and the obtained sample was fixed in 4% formalin and paraffinembedded. ACL cross-sections were stained by haematoxylin-eosin and Gallego staining for elastic and collagen fibers, and Sevier-Munger silver staining for nervous tissue. Results. ACL samples histologic degeneration classification reported 15.4% normal, 23.1% slight, 26.9% mild, 11.5% moderate and 23.1% marked. We noted 46.2% large nervous fascicles, 15.4% medium fascicles, 3.8% small fascicles, and no nerve fibers were found in 34.6% ACL samples. No significant correlation was found between the histologic degeneration and the nervous fiber quantification (p>0.05, in all cases). We noted a significant histologic degeneration inverse correlation with the VAS scale (p=0.016), and nervous fiber quantification correlation with Lequesne maximum distance walked punctuation (p=0.043). We also noted greater nervous fiber quantification with minor radiological knee osteoarthritis (Kellgren-Lawrence grade II). Conclusions. ACL degeneration and innervation deficit may play a role in primary knee osteoarthritis onset, but the lack of a defining relationship among the different parameters assessed justifies further research in greater populations

Human Bone Marrow Mesenchymal Stromal Cells Promote Bone Regeneration in a Xenogeneic Rabbit Model: A Preclinical Study

Significant research efforts have been undertaken during the last decades to treat musculoskeletal disorders and improve patient’s mobility and quality of life. The goal is the return of function as quickly and completely as possible. Cellular therapy has been increasingly employed in this setting. The design of this study was focused on cell-based alternatives. The present study aimed at investigating the bone regeneration capacity of xenogeneic human bone marrow-derived mesenchymal stromal cell (hMSC) implantation with tricalcium phosphate (TCP) granules in an immunocompetent rabbit model of critical-size bone defects at the femoral condyles. Two experimental groups, TCP and hMSC + TCP, were compared. Combination of TCP and hMSC did not affect cell viability or osteogenic differentiation. We also observed significantly higher bone regeneration in vivo in the hMSC + TCP group, which also displayed better TCP osteointegration. Also, evidence of hMSC contribution to a better TCP osteointegration was noticed. Finally, no inflammatory reaction was detected, besides the xenotransplantation of human cells into an immunocompetent recipient. In summary, hMSC combined with TCP granules is a potential combination for bone regeneration purposes that provides better preclinical results compared to TCP alone

Fourteen years of continuous soil moisture records from plant and biocrust-dominated microsites

Drylands cover ~41% of the terrestrial surface. In these water-limited ecosystems, soil moisture contributes to multiple hydrological processes and is a crucial determinant of the activity and performance of above- and belowground organisms and of the ecosystem processes that rely on them. Thus, an accurate characterisation of the temporal dynamics of soil moisture is critical to improve our understanding of how dryland ecosystems function and are responding to ongoing climate change. Furthermore, it may help improve climatic forecasts and drought monitoring. Here we present the MOISCRUST dataset, a long-term (2006–2020) soil moisture dataset at a sub-daily resolution from five different microsites (vascular plants and biocrusts) in a Mediterranean semiarid dryland located in Central Spain. MOISCRUST is a unique dataset for improving our understanding on how both vascular plants and biocrusts determine soil water dynamics in drylands, and thus to better assess their hydrological impacts and responses to ongoing climate change.This research was funded by the European Research Council (ERC Grant agreement 647038 [BIODESERT]) and Generalitat Valenciana (CIDEGENT/2018/041)

Bcmimp1, a Botrytis cinerea gene transiently expressed in planta, encodes a mitochondrial protein

<p>Botrytis cinerea is a widespread necrotrophic fungus which infects more than 200 plant species. In an attempt to characterize the physiological status of the fungus in planta and to identify genetic factors contributing to its ability to infect the host cells, a differential gene expression analysis during the interaction B. cinerea-tomato was carried out. Gene Bcmimp1 codes for a mRNA detected by differential display in the course of this analysis. During the interaction with the host, it shows a transient expression pattern with maximal expression levels during the colonization and maceration of the infected tissues. Bioinformatic analysis suggested that BCMIMP1 is an integral membrane protein located in the mitochondrial inner membrane. Co-localization experiments with a BCMIMP1-GFP fusion protein confirmed that the protein is targeted to the mitochondria. ΔBcmimp1 mutants do not show obvious phenotypic differences during saprophytic growth and their infection ability was unaltered as compared to the wild-type. Interestingly, the mutants produced increased levels of reactive oxygen species, likely as a consequence of disturbed mitochondrial function. Although Bcmimp1 expression is enhanced in planta it cannot be considered a pathogenicity factor.</p

Analysis of FOXP3 gene in children with allergy and autoimmune diseases

Allergy and autoimmunity are important immunological entities underlying chronic diseases in children. In some cases both entities develop simultaneously in the same patient. FOXP3 gene codes for a transcription factor involved in regulation of the immune system. Considering that regulatory T cells are involved in controlling immunological disease development, and the relevant role of FOXP3 in this kind of T cells, the objective of this study was to analyse the FOXP3 gene in the most prevalent autoimmune diseases and/or allergies in childhood in a European population. A total of 255 Caucasian individuals, 95 controls and 160 patients diagnosed with allergic, autoimmune or both diseases were included in this study. The molecular analysis of FOXP3 was performed by DNA sequencing following the recommendations for quality of the European Molecular Genetics Quality Network. Genomic DNA was extracted from peripheral blood of all participants and was amplified using the polymerase chain reaction. After the visualisation of the amplified fragments by agarose gel-electrophoresis, they were sequenced. Thirteen different polymorphisms in FOXP3 gene were found, seven of which had not been previously described. The mutated allele of SNP 7340C>T was observed more frequently in the group of male children suffering from both allergic and autoimmune diseases simultaneously (p=0.004, OR=16.2 [1.34-195.15]). In this study we identified for first time genetic variants of FOXP3 that are significantly more frequent in children who share allergic and autoimmune diseases. These variants mainly affect regulatory sequences that could alter the expression levels of FOXP3 modifying its function including its role in Treg cells

Filaggrin gene mutations and new SNPs in asthmatic patients: a cross-sectional study in a Spanish population

BACKGROUND: Several null-mutations in the FLG gene that produce a decrease or absence of filaggrin in the skin and predispose to atopic dermatitis and ichthyosis vulgaris have been described. The relationship with asthma is less clear and may be due to the influence of atopy in patients with associated asthma. METHODS: Four hundred individuals were included, 300 patients diagnosed with asthma divided into two groups according to their phenotype (allergic and non-allergic asthma) and 100 strictly characterized controls. The coding region and flanking regions of the FLG gene were amplified by PCR. We proceeded to the characterization of potential gene variants in that region by RFLP and sequencing and analysed their association with lung function parameters, asthma control and severity, and quality of life. RESULTS: We identified two null-mutations (R501X and 2282del4), seven SNPs previously described in databases and three SNPs that had not been previously described. One of the SNP identified in this study (1741A > T) was more frequently detected in patients with non-allergic asthma, worse FVC, FEV1 and PEF values and a higher treatment step. In addition, lowered spirometric values were observed in the non-allergic group carrying any of the nonsynonymous SNPs. CONCLUSIONS: In the association study of genetic variants of the FLG gene in our population the 1741A > T polymorphism seems to be associated with non-allergic asthma