Intensive rehabilitation programme for patients with subacute stroke in an inpatient rehabilitation facility: describing a protocol of a prospective cohort study

Accident cerebrovascular; Rehabilitació; Estudi prospectiuAccidente cerebrovascular; Rehabilitación; Estudio prospectivoRehabilitation; Stroke; Prospective StudiesRehabilitation is recognised as a cornerstone of multidisciplinary stroke care. Intensity of therapy is related to functional recovery although there is high variability on the amount of time and techniques applied in therapy sessions. There is a need to better describe stroke rehabilitation protocols to develop a better understanding of current practice increasing the internal validity and generalisation of clinical trial results. The aim of this study is to describe an intensive rehabilitation programme for patients with stroke in an inpatient rehabilitation facility, measuring the amount and type of therapies (physical, occupational and speech therapy) provided and reporting functional outcomes. Methods and analysis: This will be a prospective observational cohort study of patients with subacute stroke admitted to our inpatient rehabilitation facility during 2 years. A therapy recording tool was developed in order to describe the rehabilitation interventions performed in our unit. This tool was designed using the Delphi method, literature search and collaboration with senior clinicians. Therapists will record the time spent on different activities available in our unit during specific therapy sessions. Afterwards, the total time spent in each activity, and the total rehabilitation time for all activities, will be averaged for all patients. Outcome variables were divided into three different domains: body structure and function outcomes, activity outcomes and participation outcomes and will be assessed at baseline (admission at the rehabilitation unit), at discharge from the rehabilitation unit and at 3 and 6 months after stroke. Ethics and dissemination: This study was approved by the Medical Research Committee at Hospital del Mar Research Institute (Project ID: 34/C/2017). The results of this study will be presented at national and international congress and submitted for publication in peer-reviewed journals.This Project is funded by Fundació La Marató TV

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Unraveling the mechanisms behind the packaging of RNA into exosomes

Exosomes are small extracellular vesicles of endosomal origin of about 100nm of diameter. They are used by the cells to transfer information and can contain a wide range of biomolecules. Among many others, exosomes carry coding and non-coding RNAs. The aim of this study is to unravel the mechanisms behind the exosomal encapsulation of these biomolecules. For that purpose, we have approached this question from three different points of view. First, we studied the differences between cellular and exosomal RNA in order to determine if the class, sequence and function of the RNA molecules determine their encapsulation. Then, we assessed if two RNA binding proteins (Ago2 and GW182), often localized in the site of exosome formation, regulate the loading and export of microRNAs. Finally, we investigated how the chemotherapeutic drug Epirubicin initiates a response in the cell that leads to the exosomal over export of miR-503. In the first part of this study, we have characterized the RNA content of cellular and exosomal RNA on human endothelial cells in depth. Unlike many other profiling studies already published, we have used healthy primary cells to show the basal composition of RNA sub-classes and to find the motifs significantly more present in cells or in exosomes basally. Regarding coding genes, we have shown that mRNA genes can be classified into four different groups depending on the relative export of their regions. Moreover, we proved that RNA duos derived from the same locus and microRNA-mRNA target pairs are found at almost the same ratio in both cells and exosomes. Finally, we have also proven that, even though microRNAs are by far the most studied class of RNAs in exosomes, they represent a very small amount of the total RNA content of these vesicles thus setting the basis to promote a switch in exosomal RNA research towards longer RNAs. In the second part of this thesis, we have been able to conclude that neither AGO2 nor GW182 are involved in general processes of microRNA export based on motif recognition. Finally, in the third and last part, we have identified a new mechanism regulating microRNA export. Under basal conditions, both ANXA2 and hnRNPA2B1 bind to miR-503 in a stable partnership. Epirubicin mediates the export of miR-503 by inducing the separation of hnRNPA2B1 from the complex. In a parallel mechanism, the chemotherapeutic drug induces a reduction in the expression of hnRNPA2B1 that acts as a backup of the main complex-destabilization leading to miR-503 exosomal export

Exosomes take (germinal) center stage

Efficient antibody production is a crucial step during immune responses leading to pathogen clearance and neutralization. Immune synapses, contact points between T and B lymphocytes in the presence of an antigen, are necessary to initiate the proliferation and differentiation of B cells in the germinal center. In this issue of EMBO Reports, Fernández-Messina et al [1] present evidence of microRNA transfer from T to B cells via exosomes during synapse formation and highlight the crucial role of these exosomes for germinal center formation and the efficient production of antigen-specific antibodies

Exploring the RNA landscape of endothelial exosomes

Exosomes are small extracellular vesicles of around 100 nm of diameter produced by most cell types. These vesicles carry nucleic acids, proteins, lipids, and other biomolecules and function as carriers of biological information in processes of extracellular communication. The content of exosomes is regulated by the external and internal microenvironment of the parent cell, but the intrinsic mechanisms of loading of molecules into exosomes are still not completely elucidated. In this study, by the use of next-generation sequencing we have characterized in depth the RNA composition of healthy endothelial cells and exosomes and provided an accurate profile of the different coding and noncoding RNA species found per compartment. We have also discovered a set of unique genes preferentially included (or excluded) into vesicles. Moreover, after studying the enrichment of RNA motifs in the genes unequally distributed between cells and exosomes, we have detected a set of enriched sequences for several classes of RNA. In conclusion, our results provide the basis for studying the involvement of RNA-binding proteins capable of recognizing RNA sequences and their role in the export of RNAs into exosomes.</jats:p

Underlying the exosomal export of the microRNA, miR-503, and its implication in the resistance to chemotherapy

The communication between cancer cells and their microenvironment is an essential aspect of cancerology. Therefore, understanding how the environment could affect the tumor cells behavior is critical for the development of new anti-cancer therapies. In this study, we focus on the communication between the endothelium and breast cancer cells during neoadjuvant chemotherapy by extracellular vesicles (EVs). Extracellular vesicles (EV) are small vesicles released by cells in the extracellular milieu that mediate cell-cell communication. In the context of cancer, EVs are potentially secreted by all cell types that compose in tumor microenvironment. Previously, we identified miR-503 which exhibited downregulated or upregulated levels in exosomes released from endothelial cells under tumor environment or neoadjuvant chemotherapy, respectively. Moreover, this miRNA could affect the proliferative and invasive capacity of the tumor cells. In this study, they highlighted an exosome-dependent transfer of microRNAs from endothelial to tumor cells. This suggests that there might be a specific mechanism that sorts this microRNA into extracellular vesicles.EVs. The aim of this study is to identify the proteins involved in the export of miR-503, its impact on tumor cells behavior and how epirubicin, a chimitotherapeutic chemiotherapeutic agent, could affect it. In order to identify determine the partners of miR-503, we pulled-down a biotinylated form of thisthe microRNA and we determined the proteins attached to miR-503 using mass spectrometry. Five proteins, named miR-EXO proteins, were identified: ANXA2, hnRNPA2B1, VIM, TSP-1 and FN1.Then, we investigated which miR-EXO proteins were involved in the export of miR-503. We showed that the treatment with epirubicin induce the release of hnRNPA2B1 from the complex and, thus, allow the sorting of the microRNA. We also showed that the chemotherapy affects the localization of hnRNPA2B1. In fact, epirubicin treatment induce a switch of hnRNPA2B1 from the cytoplasm to the nucleus. Furthermore, the miR-EXO proteins were knockdowned in endothelial cells and we determined the direct impact on breast cancer cells. The hnRNPA2B1 silencing decreases the proliferation of cancer cells. According to these results, hnRNPA2B1 could inhibit the export of miR-503 into extracellular vesicles and the treatment disrupts the complex by removing hnRNPA2B1. Future work will attempt to evaluate the impact of miR-503 export during tumor growth and the clinical relevance of our findings and the implication for breast cancer patients.Mécanisme d'export des microARN dans les exosomes et de ses implications dans la réponse à la chimiothérapi

Underlying the exosomal export of a specific microRNA, miR-503, and its implication in resistance to chemotherapy

The communication between cancer cells and their microenvironment is an essential aspect of cancerology. Therefore, understanding how the environment could affect the tumor cells behavior is critical for the development of new anti-cancer therapies. In this study, we focus on the communication between the endothelium and breast cancer cells during neoadjuvant chemotherapy by extracellular vesicles (EVs). Extracellular vesicles (EV) are small vesicles released by cells in the extracellular milieu that mediate cell-cell communication. In the context of cancer, EVs are potentially secreted by all cell types that compose in tumor microenvironment. Previously, they we identified miR-503 which exhibited downregulated or upregulated levels in exosomes released from endothelial cells under tumor environment or neoadjuvant chemotherapy, respectively. Moreover, this miRNA could affect the proliferative and invasive capacity of the tumor cells. In this study, they highlighted an exosome-dependent transfer of microRNAs from endothelial to tumor cells. This suggests that there might be a specific mechanism that sorts this microRNA into extracellular vesicles.EVs. The aim of this study is to identify the proteins involved in the export of miR-503, its impact on tumor cells behavior and how epirubicin, a chimitotherapeutic chemiotherapeutic agent, could affect it. In order to identify determine the partners of miR-503, we pulled-down a biotinylated form of thisthe microRNA and we determined the proteins attached to miR-503 using mass spectrometry. Five proteins, named miR-EXO proteins, were identified: ANXA2, hnRNPA2B1, VIM, TSP-1 and FN1.Then, we investigated which miR-EXO proteins were involved in the export of miR-503. We showed that the treatment with epirubicin induce the release of hnRNPA2B1 from the complex and, thus, allow the sorting of the microRNA. We also showed that the chemotherapy affects the localization of hnRNPA2B1. In fact, epirubicin treatment induce a switch of hnRNPA2B1 from the cytoplasm to the nucleus. Furthermore, the miR-EXO proteins were knockdowned in endothelial cells and we determined the direct impact on breast cancer cells. The hnRNPA2B1 silencing decreases the proliferation of cancer cells. According to these results, hnRNPA2B1 could inhibit the export of miR-503 into extracellular vesicles and the treatment disrupts the complex by removing hnRNPA2B1. Future work will attempt to evaluate the impact of miR-503 export during tumor growth and the clinical relevance of our findings and the implication for breast cancer patients.Mécanisme d'export des microARN dans les exosomes et de ses implications dans la réponse à la chimiothérapi

hnRNPA2B1 inhibits the exosomal export of miR‑503 in endothelial cells

The chemotherapeutic drug epirubicin increases the exosomal export of miR-503 in endothelial cells. To understand the mechanisms behind this process, we transfected endothelial cells with miR-503 carrying a biotin tag. Then, we pulled-down the proteins interacting with miR-503 and studied their role in microRNA exosomal export. A total of four different binding partners were identified by mass spectrometry and validated by western blotting and negative controls, among them ANXA2 and hnRNPA2B1. Using knock-down systems combined with pull-down analysis, we determined that epirubicin mediates the export of miR-503 by disrupting the interaction between hnRNPA2B1 and miR-503. Then, both ANXA2 and miR-503 are sorted into exosomes while hnRNPA2B1 is relocated into the nucleus. The combination of these processes culminates in the increased export of miR-503. These results suggest, for the first time, that RNA-binding proteins can negatively regulate the exosomal sorting of microRNAs

Intensive rehabilitation programme for patients with subacute stroke in an inpatient rehabilitation facility: describing a protocol of a prospective cohort study

Introduction: Rehabilitation is recognised as a cornerstone of multidisciplinary stroke care. Intensity of therapy is related to functional recovery although there is high variability on the amount of time and techniques applied in therapy sessions. There is a need to better describe stroke rehabilitation protocols to develop a better understanding of current practice increasing the internal validity and generalisation of clinical trial results. The aim of this study is to describe an intensive rehabilitation programme for patients with stroke in an inpatient rehabilitation facility, measuring the amount and type of therapies (physical, occupational and speech therapy) provided and reporting functional outcomes. Methods and analysis: This will be a prospective observational cohort study of patients with subacute stroke admitted to our inpatient rehabilitation facility during 2 years. A therapy recording tool was developed in order to describe the rehabilitation interventions performed in our unit. This tool was designed using the Delphi method, literature search and collaboration with senior clinicians. Therapists will record the time spent on different activities available in our unit during specific therapy sessions. Afterwards, the total time spent in each activity, and the total rehabilitation time for all activities, will be averaged for all patients. Outcome variables were divided into three different domains: body structure and function outcomes, activity outcomes and participation outcomes and will be assessed at baseline (admission at the rehabilitation unit), at discharge from the rehabilitation unit and at 3 and 6 months after stroke. Ethics and dissemination: This study was approved by the Medical Research Committee at Hospital del Mar Research Institute (Project ID: 34/C/2017). The results of this study will be presented at national and international congress and submitted for publication in peer-reviewed journals

Genetic variants are major determinants of CSF antibody levels in multiple sclerosis

Immunological hallmarks of multiple sclerosis include the production of antibodies in the central nervous system, expressed as presence of oligoclonal bands and/or an increased immunoglobulin G index-the level of immunoglobulin G in the cerebrospinal fluid compared to serum. However, the underlying differences between oligoclonal band-positive and -negative patients with multiple sclerosis and reasons for variability in immunoglobulin G index are not known. To identify genetic factors influencing the variation in the antibody levels in the cerebrospinal fluid in multiple sclerosis, we have performed a genome-wide association screen in patients collected from nine countries for two traits, presence or absence of oligoclonal bands (n = 3026) and immunoglobulin G index levels (n = 938), followed by a replication in 3891 additional patients. We replicate previously suggested association signals for oligoclonal band status in the major histocompatibility complex region for the rs9271640*A-rs6457617*G haplotype, correlated with HLA-DRB1*1501, and rs34083746*G, correlated with HLA-DQA1*0301 (P comparing two haplotypes = 8.88 × 10(-16)). Furthermore, we identify a novel association signal of rs9807334, near the ELAC1/SMAD4 genes, for oligoclonal band status (P = 8.45 × 10(-7)). The previously reported association of the immunoglobulin heavy chain locus with immunoglobulin G index reaches strong evidence for association in this data set (P = 3.79 × 10(-37)). We identify two novel associations in the major histocompatibility complex region with immunoglobulin G index: the rs9271640*A-rs6457617*G haplotype (P = 1.59 × 10(-22)), shared with oligoclonal band status, and an additional independent effect of rs6457617*G (P = 3.68 × 10(-6)). Variants identified in this study account for up to 2-fold differences in the odds of being oligoclonal band positive and 7.75% of the variation in immunoglobulin G index. Both traits are associated with clinical features of disease such as female gender, age at onset and severity. This is the largest study population so far investigated for the genetic influence on antibody levels in the cerebrospinal fluid in multiple sclerosis, including 6950 patients. We confirm that genetic factors underlie these antibody levels and identify both the major histocompatibility complex and immunoglobulin heavy chain region as major determinants.status: publishe