Estudio, desarrollo y realización de un producto basado en sensores inerciales

Jinglob es un proyecto que resulta de la unión de música, tecnología y diseño. De la posibilidad de hacer partícipe al usuario del propio instrumento, se ha ideado un producto con el que crear música a partir de sus movimientos y gestos. El objetivo es el desarrollo de un instrumento basado en sensores inerciales que permita la creación o interpretación de música a través de una nueva forma de interacción gestual muy intuitiva, El instrumento dará respuesta a las necesidades que se derivan de un amplio rango de usuarios (tanto profesionales como aficionados, incluso aquellos sin conocimientos musicales) y de su uso en entornos muy diversos (música experimental, música en directo, actuaciones colaborativas entre músicos o diversos artistas, interacción con el público, enseñanza...

Isolation and transfection of strawverry protoplasts for gene editing

Strawberry is the most economically important soft fruit. The improvement of the organoleptic qualities of ripe fruit and the postharvest shelf life are main objectives of strawberry breeding programs. Fruit softening is mainly due to the disassembly of cell walls and the dissolution of middle lamella. In strawberry, functional analyses of genes encoding polygalacturonases (PGs) indicate that these enzymes play a key role in fruit softening, i.e. the antisense downregulation of PG genes FaPG1 or FaPG2 increased fruit firmness and postharvest shelf life (Paniagua et al., 2020). These results suggest that PG encoding genes are excellent targets for gene editing to improve strawberry fruit quality. Transfection of protoplasts with CRISPR/Cas9 ribonucleoprotein complexes is currently being explored in many species to produce DNA-free edited plants. In this research, a protocol for strawberry protoplasts transfection has been optimized with the final goal of producing non-transgenic strawberry plants with the FaPG1 gene edited. Protoplasts were isolated from 9 weeks old in vitro grown plants of Fragaria x ananassa, cv. ‘Chandler’, micropropagated in Murashige and Skoog (MS) medium supplemented with 2 mg/L of BA. Protoplast extraction and purification was performed as described by Barceló et al. (2019). Using this protocol, a yield of 1 x 105 protoplast/g fresh tissue was obtained and nearly 50-70% of them were viable. Protoplasts were transfected with the plasmid pHBT-sGFP(S65T)-NOS using a PEG-mediated transformation system, as reported by Yoo et al. (2007). To improve the efficiency of protoplast transfection, different variables were evaluated: PEG concentration, time of incubation on PEG and DNA concentration. At 48 h after transfection, the highest percentage of protoplasts showing GFP expression, 18%, was obtained with 15 minutes incubation in 20% of PEG and 5 µg of DNA

Estudio de las interacciones entre moléculas modelo de las proteínas y sales: glicina y cloruro de sodio

En este trabajo se estudió el comportamiento de los aminoácidos en disoluciones acuosas que contienen electrolitos, en concreto glicina en disolución de cloruro de sodio (NaCl).Se estableció un protocolo experimental de medida, el cual determina el método de preparación de dichas disoluciones y su conservación en el tiempo.A partir de los datos experimentales de densidades, ρ, y velocidades del sonido,<br /

Downregulation of NAC transcription factors modifies cell wall composition and increases strawberry fruit firmness

The strawberry is a soft fruit with a very short post-harvest shelf life. The changes in texture during fruit ripening are mainly due to the dissolution of the middle lamellae, reducing cell-to-cell adhesion, and the weakening of parenchymal cell walls as result of the action of cell wall modifying enzymes. At present, no master regulator of this process has been discovered yet. NAC transcription factors have been involved in numerous physiological processes, including fruit ripening. In strawberry, the NAC family comprises more than 110 genes, and at least 6 of them are expressed during fruit development. In this research, we performed a functional analysis of two ripening-related NAC genes, FaNAC2 and FaNAC3, in Fragaria x ananassa Duch. cv. Chandler. Several RNAi transgenic lines showing low FaNAC2 or FaNAC3 mRNA levels in fruit were obtained through Agrobacterium-mediated transformation. These lines produced fruits significantly firmer than control at the ripe stage, being the increase in firmness higher in FaNAC2 silenced plants. Cell walls were extracted from ripe transgenic fruits and characterized by ELISA and Epitope Detection Chromatography (EDC), using monoclonal antibodies against different polysaccharide epitopes. FaNAC2 transgenic lines showed more extensive changes than FaNAC3; these modifications involved increased amounts of demethylated pectins (LM19) in water and CDTA fractions and an alteration of the lateral branches of RG-I, decreasing the amount of arabinan epitopes and increasing galactan epitopes detected by LM6 and LM5, respectively. The amount of arabinogalactan proteins recognized by the JIM13 antibody was also affected, decreasing in the Na2CO3 fraction and increasing in the 4M KOH and cellulase fraction of the transgenic lines.The results obtained indicate that NAC genes could be involved in the regulation of cell wall disassembly associated to strawberry fruit softening.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

High-throughput mapping of cell wall glycans to unveil cell wall disassembly, a key process determining strawberry fruit softening

The short shelf life of strawberry fruit is a major limitation that produces important economic losses related to postharvest spoiling. Fruit texture of fleshy fruits is a complex trait but mainly rely on mechanical properties of parenchyma cell walls. Several studies support the relevance of cell wall modifying enzymes on cell wall deconstruction, decreasing cell wall strength and cell to cell adhesion, and ultimately producing the softening of the fruit at macroscopic level. Previous studies on our group showed that transgenic silencing of ripening-specific genes encoding some of these enzymes reduced softening and increased postharvest shelf life in strawberry (Fragaria × ananassa, cv. ‘Chandler’) fruits. In this research, to further investigate the cell wall remodelling process associated to strawberry softening a high-throughput analysis of cell wall composition based on monoclonal antibodies against different polysaccharide epitopes has been performed. To this purpose, cell walls were isolated from non-transgenic fruits at different developmental stages as well as from ripe fruits of selected transgenic lines with genes involved in metabolism of pectins (pectate lyase, polygalacturonase, β-galactosidase, pectin acetil esterase), hemicellulose/cellulose (endo-β-glucanase) or lignin (cinnamyl alcohol dehydrogenase) down-regulated. These transgenic lines showed a large variability in fruit firmness at ripening. Cell walls were fractionated and subjected to a carbohydrate microarray. The results obtained unveiled a common pattern of cell wall composition on those transgenic lines with firmer phenotypes, specially defined by the higher content of pectins on those cell wall fractions more imbricated in the matrix, which can be interpreted as a less degraded cell wall structure.This research was supported by FEDER EU Funds and the Ministerio de Economía y Competitividad of Spain (grant reference AGL2017-86531-C2-1-R). Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

External Validation of SAFE Score to Predict Atrial Fibrillation Diagnosis after Ischemic Stroke: A Retrospective Multicenter Study

Diagnóstico; Fibrilación auricular; Ictus isquémicoDiagnòstic; Fibril·lació auricular; Ictus isquèmicDiagnosis; Atrial fibrillation; Ischemic strokeIntroduction: The screening for atrial fibrillation (AF) scale (SAFE score) was recently developed to provide a prediction of the diagnosis of AF after an ischemic stroke. It includes 7 items: age ≥ 65 years, bronchopathy, thyroid disease, cortical location of stroke, intracranial large vessel occlusion, NT-ProBNP ≥250 pg/mL, and left atrial enlargement. In the internal validation, a good performance was obtained, with an AUC = 0.88 (95% CI 0.84-0.91) and sensitivity and specificity of 83% and 80%, respectively, for scores ≥ 5. The aim of this study is the external validation of the SAFE score in a multicenter cohort. Methods: A retrospective multicenter study, including consecutive patients with ischemic stroke or transient ischemic attack between 2020 and 2022 with at least 24 hours of cardiac monitoring. Patients with previous AF or AF diagnosed on admission ECG were excluded. Results: Overall, 395 patients were recruited for analysis. The SAFE score obtained an AUC = 0.822 (95% CI 0.778-0.866) with a sensitivity of 87.2%, a specificity of 65.4%, a positive predictive value of 44.1%, and a negative predictive value of 94.3% for a SAFE score ≥ 5, with no significant gender differences. Calibration analysis in the external cohort showed an absence of significant differences between the observed values and those predicted by the model (Hosmer-Lemeshow's test 0.089). Conclusions: The SAFE score showed adequate discriminative ability and calibration, so its external validation is justified. Further validations in other external cohorts or specific subpopulations of stroke patients might be required.We acknowledge FIBAO (Biomedical Research Foundation) and Adrián Aparicio Mota for their assistance with statistical analysis. Adrián Aparicio (a FIBAO statistician) analyzed the collected data. The project was the winner of the IV research grant “STROKE PROJECT 2020” from the Spanish Society of Neurology

Non-criteria obstetric antiphospholipid syndrome: how different is from Sidney criteria? A single-center study

This study aims to compare the demographic characteristics, clinical features, serology, and fetal-maternal outcomes between women with obstetric antiphospholipid syndrome (APS) and those with non-criteria (NC)-APS and seronegative (SN)-APS. Two-hundred and sixty-three women with APS obstetric morbidity ever pregnant were included. Of those, 66 met the APS classification criteria, 140 were NC-APS, and 57 were SN-APS. Patients with other autoimmune diseases were excluded. Adverse pregnancy outcomes (APO) included early pregnancy loss, fetal death, preeclampsia, abruptio placentae, and preterm birth. The mean age of the study group was 33.6 ± 5.3 years, and patients were followed up for 129.5 ± 81.9 months. In the NC-APS group, 31 (22.1%) did not fulfill clinical and serological criteria (Subgroup A), 49 (35%) did meet clinical but not serologic criteria (Subgroup B), and 60 (42.9%) fulfilled the serologic criteria but not the clinical ones (Subgroup C). The cardiovascular risk burden was higher in the APS group, due to a higher proportion of smoking. Patients with criteria APS received more intensive treatment than patients in the other study groups. The addition of standard of care (SoC) treatment significantly improved live birth and decreased APO in all groups. Significant clinical differences were observed between the study groups. However, when treated with SoC, fetal-maternal outcomes were similar, with a significant improvement in live births and a decrease in APO. Risk stratification in patients with obstetric morbidity associated with APS can help individualize their treatment

Prevalence of hospital PCR-confirmed COVID-19 cases in patients with chronic inflammatory and autoimmune rheumatic diseases

[Abstract] Background: The susceptibility of patients with rheumatic diseases and the risks or benefits of immunosuppressive therapies for COVID-19 are unknown. Methods: We performed a retrospective study with patients under follow-up in rheumatology departments from seven hospitals in Spain. We matched updated databases of rheumatology patients with severe acute respiratory syndrome coronavirus 2-positive PCR tests performed in the hospital to the same reference populations. Rates of PCR+ confirmed COVID-19 were compared among groups. Results: Patients with chronic inflammatory diseases had 1.32-fold higher prevalence of hospital PCR+ COVID-19 than the reference population (0.76% vs 0.58%). Patients with systemic autoimmune or immune-mediated disease (AI/IMID) showed a significant increase, whereas patients with inflammatory arthritis (IA) or systemic lupus erythematosus did not. COVID-19 cases in some but not all diagnostic groups had older ages than cases in the reference population. Patients with IA on targeted-synthetic or biological disease-modifying antirheumatic drugs (DMARDs), but not those on conventional-synthetic DMARDs, had a greater prevalence despite a similar age distribution. Conclusion: Patients with AI/IMID show a variable risk of hospital-diagnosed COVID-19. Interplay of ageing, therapies and disease-specific factors seem to contribute. These data provide a basis to improve preventive recommendations to rheumatic patients and to analyse the specific factors involved in COVID-19 susceptibility.Instituto de Salud Carlos III; RD16/0012 RETICS Progra

A functional variant of TLR10 modifies the activity of NFkB and may help predict a worse prognosis in patients with rheumatoid arthritis

Background: Toll-like receptor (TLR) family members are key players in inflammation. TLR10 has been poorly studied in chronic inflammatory disorders, and its clinical relevance in rheumatoid arthritis (RA) is as yet unknown. We aimed at identifying TLR10 variants within all coding regions of the gene in patients with RA as well as studying their functional and clinical significance. Methods: TLR10 gene variants were studied by performing sequencing of 66 patients with RA and 30 control subjects. A selected variant, I473T, was then analyzed in 1654 patients and 1702 healthy control subjects. The capacity of this TLR10 variant to modify the transcriptional activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB) was determined by using a luciferase reporter assay and analyzing the expression of NFkB target genes by quantitative polymerase chain reaction. Differences between groups were analyzed by using the Mann-Whitney U test and the unpaired two-tailed Student’s t test. Results: We detected ten missense variants in the TLR10 gene and focused on the I473T substitution based on allele frequencies and the predicted functional impact. I473T variant is not associated with susceptibility to RA, but it significantly correlates with erosive disease in patients seropositive for antibodies to citrullinated protein antigens (p = 0.017 in the total cohort and p = 0.0049 in female patients) and with a lower response to infliximab treatment as measured by the change in Disease Activity Score in 28 joints (p = 0.012) and by the European League Against Rheumatism criteria (p = 0.049). Functional studies showed that TLR10 reduced activation of the NFkB inflammatory pathway in hematopoietic cells, whereas the I473T variant lacked this inhibitory capacity. Consistently, after exposure to infliximab, cells expressing the I437T variant showed higher NFkB activity than cells carrying wild-type TLR10. Conclusions: A TLR10 allelic variant, I473T, has impaired NFkB inhibitory activity and is highly associated with disease severity and low response to infliximab in patients with RA

Mitochondrial DNA haplogroups influence the risk of incident knee osteoarthritis in OAI and CHECK cohorts: a meta-analysis and functional study

Extended report[Abstract] Objective To evaluate the influence of the mitochondrial DNA (mtDNA) haplogroups in the risk of incident knee osteoarthritis (OA) and to explain the functional consequences of this association to identify potential diagnostic biomarkers and therapeutic targets. Methods Two prospective cohorts contributed participants. The osteoarthritis initiative (OAI) included 2579 subjects of the incidence subcohort, and the cohort hip and cohort knee (CHECK) included 635, both with 8-year follow-up. The analysis included the association of mtDNA haplogroups with the rate of incident knee OA in subjects from both cohorts followed by a subsequent meta-analysis. Transmitochondrial cybrids harbouring haplogroup J or H were constructed to detect differences between them in relation to physiological features including specific mitochondrial metabolic parameters, reactive oxygen species production, oxidative stress and apoptosis. Results Compared with H, the haplogroup J associates with decreased risk of incident knee OA in subjects from OAI (HR=0.680; 95% CI 0.470 to 0.968; p<0.05) and CHECK (HR=0.728; 95% CI 0.469 to 0.998; p<0.05). The subsequent meta-analysis including 3214 cases showed that the haplogroup J associates with a lower risk of incident knee OA (HR=0.702; 95% CI 0.541 to 0.912; p=0.008). J cybrids show a lower free radical production, higher cell survival under oxidative stress conditions, lower grade of apoptosis as well as lower expression of the mitochondrially related pro-apoptotic gene BCL2 binding component 3 (BBC3). In addition, J cybrids also show a lower mitochondrial respiration and glycolysis leading to decreased ATP production. Conclusions The physiological effects of the haplogroup J are beneficial to have a lower rate of incident knee OA over time. Potential drugs to treat OA could focus on emulating the mitochondrial behaviour of this haplogroup.Instituto de Salud Carlos III; CIBERCB06/01/0040-SpainInstituto de Salud Carlos III; RETIC-RIER-RD12/0009/0018Instituto de Salud Carlos III; PI14/01254Instituto de Salud Carlos III; CP12/0319