Publicaciones

The proteomic signature of circulating extracellular vesicles following intracerebral hemorrhage: Novel insights into mechanisms underlying recovery

Casado-Fernández L,Laso-García F, Piniella D et al.  Neurobiol Dis. 2024 Oct 15;201:106665. doi: 10.1016/j.nbd.2024.106665. Epub 2024 Sep 12. PMID: 39277144.

https://pubmed.ncbi.nlm.nih.gov/39277144/

Abstract: Circulating extracellular vesicles (EVs) can participate in innate repair processes triggered after intracerebral hemorrhage (ICH). We aimed to describe changes in the proteomic profile of circulating EVs between the acute and subacute phases of ICH and to compare the findings depending on outcomes, as an approach to unraveling such repair mechanisms. This was a prospective observational study including patients with non-traumatic supratentorial ICH. Exclusion criteria were previous disability, signs of herniation on baseline computed tomography, or limited life expectancy. EVs were isolated from blood samples at 24 h and 7 days after symptom onset. After 6-months’ follow- up, patients were dichotomized into poor and good outcomes, defining good as an improvement of >10 points or >50 % on the National Institutes of Health Stroke Scale and a modified Rankin Scale of 0–2. The protein cargo was analyzed by quantitative mass spectrometry and compared according to outcomes. Forty-four patients completed follow-up, 16 (35.5 %) having good outcomes. We identified 1321 proteins in EVs, 37 with differential abundance. In patients with good outcomes, proteins related to stress response (DERA, VNN2, TOMM34) and angiogenesis (RHG01) had increased abundance at 7 days. EVs from patients with poor outcomes showed higher levels of acute-phase reactants (CRP, SAA2) at 7 days compared with 24 h. In conclusion, the protein content of circulating EVs in patients with ICH changes over time, the changes varying depending on the clinical outcome, with greater abundance of proteins potentially involved in the repair processes of patients with good outcomes.

Funding: This work was supported by Carlos III Health Institute (ISCIII) and co-funded by the European Union (European Regional Development Fund-FEDER) under grant PI17/01922 and PI20/00243, Invictus plus network under grant RD16/0019/0005, RICORS ICTUS network under grant RD21/0006/0012 and the Next Generation EU fundig for actions in the Recovery and Resilience Mechanism, Miguel Servet under grant CPII20/00002 to MG-F; CP20/00024 to LO-O, Ministerio de Universidades, Plan de Recuperaci´ on, Transformaci´ on y Resiliencia y la Universidad Aut´ onoma de Madrid under grant CA1/RSUE/2021-00753 to DP, and the Spanish Ministry of Health- (ISCIII) under grant CM20/ 00047 to EA-L, CM23/00022 to LC-F, FI18/00026 to FL-G and FI17/ 00188 to MG-dF. We also thank the editing assistance of Morote Traducciones.

Dubbing language-therapy CINEma-based in aphasia post-stroke (DULCINEA): A feasibility randomized crossover controlled trial

Fuentes B, Jordi-Perea P, Sempere-Iborra C et al. Digit Health. 2024 Oct 10;10:20552076241288311. doi: 10.1177/20552076241288311. PMID: 39421311.

https://pubmed.ncbi.nlm.nih.gov/39421311/

Abstract: Background: Helping people recover from aphasia is among the top 10 research priorities relating to life after stroke. Objective: We aimed to evaluate the feasibility of dubbing techniques (using newly developed software) for post-stroke aphasia therapy and explore its potential efficacy. Methods: Randomised, crossover, interventional, feasibility trial that included patients with chronic post-stroke non-fluent aphasia. The intervention consisted of an individualised programme (16 sessions; 8 weeks) based on dubbing words and sentences progressively adapted to the severity of the aphasia. Patients were allocated to groups that underwent therapy within the first 3 months, or between 3 and 6 months from inclusion, each group serving as the control during the nontherapy periods. Outcomes were the pre-post differences in the Communicative Activity Log, the Boston Diagnostic Aphasia Examination, the General Health Questionnaire-12, the Stroke Aphasia Quality of Life Scale, and the Western Aphasia Battery Revised, administered by psychologists blinded to the patients’ allocation. Results: Recruitment was limited due to COVID-19 and prematurely stopped because of funding coming to an end. A total of 23 patients were randomised, 20 of whom completed the study (1 withdrew consent, and 2 dropped out). The adherence rate to the allocated group was 95.3%. No statistically significant differences were found in any of the outcomes; however, 17 (85%) patients reported subjective improvements in communication skills. Conclusions: This trial shows the feasibility of dubbing therapy (using dedicated software) for patients with post-stroke nonf luent aphasia. Although it lacks statistical power, certain effects on language and communication cannot be ignored.

Funding: This study was promoted by the Research Foundation of La Paz University Hospital, which hosted a research consortium joined by the Department of Neurology at La Paz University Hospital, the Department of Psychology at Comillas Pontifical University, and the patients’ association, Afasia Activa. This project has received funding from “la Caixa” Banking Foundation under the project code LCF/PR/HR19/52160009. The funder was not involved in any of the following processes: design of the trial, data collection, analysis, or interpretation, or writing the manuscript. BF, EdC-R, RR, GR-A, JR-P, EA and MA-L are members of the Spanish Stroke Research Network RICORS ICTUS (RD21/0006/0012) funded by the Carlos III Institute of Health and the European Union (NextGenerationEU). ‘la Caixa’ Foundation, Instituto de Salud Carlos III, (grant number LCF/PR/ HR19/52160009, RD21/0006/0012).

 

Mediterranean Diet Prior to Ischemic Stroke and Potential Circulating Mediators of Favorable Outcomes

María Castañón-Apilánez, Carmen García-Cabo, Cristina Martin-Martin et al. Nutrients. 2024 Sep 23;16(18):3218.doi: 10.3390/nu16183218.. PMID: 39339817

https://pubmed.ncbi.nlm.nih.gov/39339817/

Abstract: Background/Objectives. A Mediterranean diet (MD) has been associated with neuroprotective effects. We aimed to assess the MD’s association with stroke prognosis and the potential mediators involved. Methods. Seventy patients with acute anterior circulation ischemic stroke were included. Dietary patterns were evaluated using the MEDAS scale, a food-frequency questionnaire, and a 24 h recall. Circulating biomarkers including insulin resistance (HOMA index), adipokines (resistin, adiponectin, leptin), choline pathway metabolites (TMAO, betaine, choline), and endothelial progenitor cells (EPCs) were measured. Early neurological improvement (ENI) at 24 h, final infarct volume, and functional outcome at 3 months were assessed. Results. Adherence to MD and olive oil consumption were associated with a lower prevalence of diabetes and atherothrombotic stroke, and with lower levels of fasting glycemia, hemoglobinA1C, insulin resistance, and TMAO levels. Monounsaturated fatty acids and oleic acid consumption correlated with lower resistin levels, while olive oil consumption was significantly associated with EPC mobilization. Multivariate analysis showed that higher MD adherence was independently associated with ENI and good functional prognosis at 3 months. EPC mobilization, lower HOMA levels, and lower resistin levels were associated with ENI, a smaller infarct volume, and good functional outcome. Conclusions. MD was associated with better prognosis after ischemic stroke, potentially mediated by lower insulin resistance, increased EPC mobilization, and lower resistin levels, among other factors.

Funding: This study has been funded by Instituto de Salud Carlos III (ISCIII) through RICORS-ICTUS (RD21/0006/0022, PI18/01096, CM19/00235, INT19/00075) by means of NextGenerationEU funds, which support the actions of the Mecanismo de Recuper-ación y Resiliencia (MRR)/PRTR 

 

The mitochondrial Na+/Ca2+ exchanger NCLX is implied in the activation of hypoxia-inducible factors

Carmen Choya-Foces , Elisa Navarro , Cristóbal de Los Ríos et al. Redox Biol. 2024 Nov:77:103364. doi: 10.1016/j.redox.2024.103364. PMID: 39341036.

https://pubmed.ncbi.nlm.nih.gov/39341036/

Abstract: Eukaryotic cells and organisms depend on oxygen for basic living functions, and they display a panoply of adaptations to situations in which oxygen availability is diminished (hypoxia). A number of these responses in animals are mediated by changes in gene expression programs directed by hypoxia-inducible factors (HIFs), whose main mechanism of stabilization and functional activation in response to decreased cytosolic oxygen concentration was elucidated two decades ago. Human acute responses to hypoxia have been known for decades, although their precise molecular mechanism for oxygen sensing is not fully understood. It is already known that a redox component, linked with reactive oxygen species (ROS) production of mitochondrial origin, is implied in these responses. We have recently described a mechanism by which the mitochondrial sodium/calcium exchanger, NCLX, participates in mitochondrial electron transport chain regulation and ROS production in response to acute hypoxia. Here we show that NCLX is also implied in the response to hypoxia mediated by the HIFs. By using a NCLX inhibitor and interference RNA we show that NCLX activity is necessary for HIF-α subunits stabilization in hypoxia and for HIF-1-dependent transcriptional activity. We also show that hypoxic mitochondrial ROS production is not required for HIF-1α stabilization under all circumstances, suggesting that the basal cytosolic redox state or other mechanism(s) could be operating in the NCLX-mediated response to hypoxia that operates through HIF-α stabilization. This finding provides a link between acute and medium-term responses to hypoxia, reinforcing a central role of mitochondrial cell signalling in the response to hypoxia.

Funding: This research has been financed by grants from the Spanish Government (partially funded by the European Union ERDF “A way of making Europe” and Next GenerationEU): RedoxStroke (RTI2018- 094203-B-I00), excellence network RED2018-102576-T, NCLRedoX (PID2021-124688OB-I00), NCLXtroke (PDC2022-133246-I00),
PID2021-125986OB-I00 and PID2022-139936OA-I00 from AEI (MICIU/ AEI/10.13039/501100011033); PI15/00107, PI22/00362 and RICORSICTUS (RD21/0006/0009) from Instituto de Salud Carlos III (ISCIII) and fellowships IJC2020-042679-I (MICIU) and RYC2022-036516-I (MICIU) to P.H.-A. from AEI and FPU18/03475 to C.C.-F. from MICIU, and by a grant from the Fundacion ´ Domingo Martínez.

A dual colorimetric-electrochemical microfluidic paper-based analytical device for point-of-care testing of ischemic strokes

Dortez S, Pacheco M, Gasull T, Crevillen AG, Escarpa A. Lab Chip. 2024 Aug 9. doi: 10.1039/d4lc00398e. Online ahead of print. PMID: 39118539
Abstract: A novel microfluidic paper-based analytical device with dual colorimetric and electrochemical detection (dual μPAD) was developed for the assessment of transferrin saturation (TSAT) in samples from ischemic stroke patients. TSAT was calculated from the ratio between transferrin-bound iron, which was colorimetrically measured, and the total iron-binding capacity, which was electrochemically measured. To this end, a μPAD was smartly designed, which integrated both colorimetric and electrochemical detection reservoirs, communicating via a microchannel acting as a chemical reactor, and with preloading/storing capabilities (reagent-free device). This approach allowed the dual and simultaneous determination of both parameters, providing an improvement in the reliability of the results due to an independent signal principle and processing. The μPADs were validated by analyzing a certified reference material, showing excellent accuracy (Er ≤ 5%) and precision (RSD ≤ 2%). Then they were applied to the analysis of diagnosed serum samples from ischemic stroke patients. The results were compared to those provided by a free-interference method (urea-PAGE). Impressively, both methods exhibited a good correlation (r = 0.96, p < 0.05) and no significant differences were found between them (slope 1.0 ± 0.1 and the intercept 1 ± 4, p < 0.05), demonstrating the excellent accuracy of our approach during the analysis of complex samples from ischemic stroke patients, using just 90 μL of clinical samples and taking less than 90 min in comparison with the 18 hours required by the urea-PAGE approach. The developed fully integrated colorimetric-electrochemical μPAD is a promising ready to use reagent-free device for the point-of-care testing of TSAT, which can be used to assist physicians in the fast diagnosis and prognosis of ischemic strokes, where the decision-time is crucial for the patient’s survival.
Funding: This work has been financially supported by the TRANSNANOAVANSENS program from the Community of Madrid (P2018/NMT-4349) (A. E.), by the grant PID2020-118154GB-I00 funded by MCIN/AEI/10.13039/501100011033 (A. E.), by the NEURO-CHIP-CM program from the Community of Madrid (Y2020/NMT6312) (A. E.), by the RICORS RD21/0006/0024 (NextGeneration EU funding) and 2021SGR00925 (Agencia de gestio d’Ajuts Universitaris i de Recerca de Catalunya) (T. G.), and by the Spanish Ministry of Economy and Competitiveness (CTQ2017-86441-C2-1-R, FPI fellowship (S. D.)).

Atrial Imaging and Cardiac Rhythm in Cryptogenic Embolic Stroke: The ARIES Study

Rigual R,Castrejón-Castrejón S, Fernández-Gassó L et al. J Am Heart Assoc. 2024 Sep 3;13(17):e036236. doi: 10.1161/JAHA.124.036236. Epub 2024 Aug 29. PMID: 39206739.

https://pubmed.ncbi.nlm.nih.gov/39206739/

Abstract: BACKGROUND: Unknown cardioembolic sources are frequent causes of cryptogenic stroke. We analyzed the risk of atrial fibrillation (AF) or high burden of ectopic atrial activity (HBEA) in patients with cryptogenic stroke, assessing atrial function and 1- year outcomes. METHODS AND RESULTS: The ARIES (Atrial Imaging and Cardiac Rhythm in Cryptogenic Embolic Stroke) study is an observational study including patients with cryptogenic stroke. We analyzed the frequency of AF and HBEA (>3000 atrial ectopic beats/day or >2 bursts or atrial tachycardia between 3 beats and ≤30 seconds) in two 30- day Holter- ECGs, comparing advanced echocardiography signs of left atrial (LA) dysfunction according to rhythm: AF, HBEA, and normal sinus rhythm. We also evaluated 1- year stroke recurrence and mortality. The study included 109 patients; 35 (32.1%) patients had AF, 27 (24.8%) HBEA, and 47 (43.1%) normal sinus rhythm. Compared with those with normal sinus rhythm, patients with AF presented higher 2- dimensional and 3- dimensional LA indexed volumes (38.8±11.2 versus 27.3±11.8 mL/m2, and 50.6±17.2 versus 34.0±15.4 mL/m2, respectively, P<0.001), lower 3- dimensional LA ejection fraction (50±14.6 versus 62.7±11.8, P=0.001), LA reservoir strain (22.0±8.6 versus 30.4±10.5, P<0.001), and LA contraction strain (10.5±8.18 versus 17.1±7.5, P<0.001), remaining significant in multivariate analysis. Patients with HBEA showed higher LA indexed volumes and lower LA reservoir strain than patients with normal sinus rhythm only in univariate analysis. There were no differences in ischemic recurrence or mortality among the groups. CONCLUSIONS: Patients with cryptogenic stroke showed a high incidence of AF and HBEA. AF is strongly related to LA volume, LA function, and LA reservoir and contraction strain, whereas HBEA showed milder structural changes. Advanced LA echocardiography could help patient selection for long- term ECG monitoring in suspected cardiac sources.

Funding: This project has been supported by a grant from the Stroke Project of the Cerebrovascular Diseases Study Group of the Spanish Society of Neurology, funded by Daiichi- Sankyo. This work was also supported by RICORS (Redes de Investigación Cooperativa Orientadas a Resultados en Salud) network under grant RD21/0006/0012.