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Introduction: Magnetic resonance imaging (MRI) is crucial for diagnosing and monitoring of multiple sclerosis (MS) as it is used to assess lesions in the brain and spinal cord. However, in real-world clinical settings, MRI scans are often acquired with thick slices, limiting their utility for automated quantitative analyses. This work presents a single-image super-resolution (SR) reconstruction framework that leverages SR convolutional neural networks (CNN) to enhance the through-plane resolution of structural MRI in people with MS (PwMS)., Methods: Our strategy involves the supervised fine-tuning of CNN architectures, guided by a content loss function that promotes perceptual quality, as well as reconstruction accuracy, to recover high-level image features., Results: Extensive evaluation with MRI data of PwMS shows that our SR strategy leads to more accurate MRI reconstructions than competing methods. Furthermore, it improves lesion segmentation on low-resolution MRI, approaching the performance achievable with high-resolution images., Discussion: Results demonstrate the potential of our SR framework to facilitate the use of low-resolution retrospective MRI from real-world clinical settings to investigate quantitative image-based biomarkers of MS., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Giraldo, Khan, Pineda, Liang, Lozano-Castillo, Van Wijmeersch, Woodruff, Lambin, Romero, Peeters and Sijbers.)

This paper explores the significant role of real-world data (RWD) in advancing our understanding and management of Multiple Sclerosis (MS). RWD has proven invaluable in MS research and care, offering insights from larger and diverse patient populations. A key focus of the paper is the European Health Data Space (EHDS), a significant development that promises to change how healthcare data is managed across Europe. This initiative is particularly relevant to the MS community. The paper highlights various data initiatives, discussing their importance for those affected by MS. Despite the potential benefits, there are challenges and concerns, especially about ensuring that the growth of various data platforms remains beneficial for MS patients. The paper suggests practical actions for the global MS community to consider, aimed at optimizing the use of RWD. The emphasis of this discussion is on the secondary use of health data, particularly in the European context. The content is based on the author's own experiences and interpretations, offering a personal yet informed view on using RWD to improve MS research and patient care., Competing Interests: The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Peeters.)

Real-world data (RWD) has the potential to revolutionize healthcare by offering valuable insights into patient outcomes and treatment efficacy. However, leveraging RWD effectively presents challenges, including its inherent limitations, diverse stakeholders, and insufficient data management pipelines. A proposed framework advocates three essential elements: adherence to FAIR principles (Findable, Accessible, Interoperable, and Reusable), stakeholder engagement and education, and highlighting the need for inclusive, pragmatic federated hybrid pipelines. By employing these strategies, healthcare organizations can overcome obstacles to RWD utilization and foster sustainable progress in patient care.

Background: The integrity and reliability of clinical research outcomes rely heavily on access to vast amounts of data. However, the fragmented distribution of these data across multiple institutions, along with ethical and regulatory barriers, presents significant challenges to accessing relevant data. While federated learning offers a promising solution to leverage insights from fragmented data sets, its adoption faces hurdles due to implementation complexities, scalability issues, and inclusivity challenges., Objective: This paper introduces Federated Learning for Everyone (FL4E), an accessible framework facilitating multistakeholder collaboration in clinical research. It focuses on simplifying federated learning through an innovative ecosystem-based approach., Methods: The "degree of federation" is a fundamental concept of FL4E, allowing for flexible integration of federated and centralized learning models. This feature provides a customizable solution by enabling users to choose the level of data decentralization based on specific health care settings or project needs, making federated learning more adaptable and efficient. By using an ecosystem-based collaborative learning strategy, FL4E encourages a comprehensive platform for managing real-world data, enhancing collaboration and knowledge sharing among its stakeholders., Results: Evaluating FL4E's effectiveness using real-world health care data sets has highlighted its ecosystem-oriented and inclusive design. By applying hybrid models to 2 distinct analytical tasks-classification and survival analysis-within real-world settings, we have effectively measured the "degree of federation" across various contexts. These evaluations show that FL4E's hybrid models not only match the performance of fully federated models but also avoid the substantial overhead usually linked with these models. Achieving this balance greatly enhances collaborative initiatives and broadens the scope of analytical possibilities within the ecosystem., Conclusions: FL4E represents a significant step forward in collaborative clinical research by merging the benefits of centralized and federated learning. Its modular ecosystem-based design and the "degree of federation" feature make it an inclusive, customizable framework suitable for a wide array of clinical research scenarios, promising to revolutionize the field through improved collaboration and data use. Detailed implementation and analyses are available on the associated GitHub repository., (©Ashkan Pirmani, Martijn Oldenhof, Liesbet M Peeters, Edward De Brouwer, Yves Moreau. Originally published in JMIR Formative Research (https://formative.jmir.org), 17.07.2024.)

Introduction: The primary objective of the core data set is to reduce heterogeneity and promote harmonization among data sources in EM, thereby reducing the time needed to execute real life data collection efforts. Recently, a group led by the Multiple Sclerosis Data Alliance has developed a core data set for collecting real-world data on multiple sclerosis (MS) globally. Our objective was to adapt this global data set to the needs of Latin America, so that it can be implemented by the registries already developed and in the process of development in the region., Material and Methods: A working group was formed regionally, the core data set created globally was adapted (translation process into Spanish, incorporation of regional variables and consensus on variables to be used). Consensus was obtained through the remote Delphi methodology of a round of questionnaires and remote discussion of the core data set variables., Results: A total of 25 professionals from Latin America carried out the adaptation process between November 2022 and July 2023. Agreement was established on a core data set of nine categories and 45 variables, version 2023 to suggest its implementation in developed or developing registries, and MS cohorts in the region., Conclusion: The core data set seeks to harmonize the variables collected by registries and cohorts in MS in Latin America in order to facilitate said collection and allow collaboration between sources. Its implementation will facilitate real life data collection and collaboration in the region.

Background: As of September 2022, there was no globally recommended set of core data elements for use in multiple sclerosis (MS) healthcare and research. As a result, data harmonisation across observational data sources and scientific collaboration is limited., Objectives: To define and agree upon a core dataset for real-world data (RWD) in MS from observational registries and cohorts., Methods: A three-phase process approach was conducted combining a landscaping exercise with dedicated discussions within a global multi-stakeholder task force consisting of 20 experts in the field of MS and its RWD to define the Core Dataset., Results: A core dataset for MS consisting of 44 variables in eight categories was translated into a data dictionary that has been published and disseminated for emerging and existing registries and cohorts to use. Categories include variables on demographics and comorbidities (patient-specific data), disease history, disease status, relapses, magnetic resonance imaging (MRI) and treatment data (disease-specific data)., Conclusion: The MS Data Alliance Core Dataset guides emerging registries in their dataset definitions and speeds up and supports harmonisation across registries and initiatives. The straight-forward, time-efficient process using a dedicated global multi-stakeholder task force has proven to be effective to define a concise core dataset., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: T.P. has no other conflicts of interests to disclose than that she is funded by the Flemish Government under the ‘Special Research Fund (Bijzonder Onderzoeksfonds, BOF)’: BOF22OWB01. L.G. has no other conflicts of interests to disclose than that she is funded by the Flemish Government under the ‘Onderzoeksprogramma Artificiële Intelligentie Vlaanderen’. A.H. has no personal pecuniary interests to disclose, other than being an employee of the MS International Federation, which receives income from a range of corporate sponsors, recently including: Biogen, BristolMyersSquibb, Coloplast, Janssen, Merck, Viatris (formerly Mylan), Novartis, Roche and Sanofi – all of which is publicly disclosed. J.H. has received honoraria for serving on advisory boards for Biogen, Bristol-Myers-Squibb, Sanofi-Genzyme, Merck KGaA, Novartis, and Sandoz and speaker’s fees from Biogen, Novartis, Merck KGaA, Teva and Sanofi-Genzyme. He has served as P.I. for projects, or received unrestricted research support from, Biogen, Bristol-Myers-Squibb, Merck KGaA, Novartis, Roche and Sanofi-Genzyme. His MS research was funded by the Swedish Research Council and the Swedish Brain foundation. H.S. works for the Accelerated Cure Project for MS (ACP), which has received grants, collaboration funding, payments for use of assets, or in-kind contributions from the following companies: EMD Serono, Sanofi/Genzyme, Biogen, Genentech, AbbVie, Octave, GlycoMinds, Pfizer, MedDay, AstraZeneca, Teva, Mallinckrodt, MSDx, Regeneron Genetics Centre, BC Platforms, and Celgene. A.C.P. has also received funding from the Patient-Centred Outcomes Research Institute (PCORI) and the National MS Society (NMSS). A.S. is on the editorial board for Neurology and serves as a consultant for Gryphon Bio, LLC. She has received research funding from the Department of Defence, MS Society of Canada, National MS Society and the Consortium of MS Centres. A.S. works for the NARCOMS Registry which is supported by the Consortium of MS Centres (CMSC) and the Foundation of the CMSC. R.M. has received no personal funding from any sources, the UK MS Register is funded by the MS Society, and has received funding for specific projects from Novartis, Sanofi-Genzyme, and Merck KGaA. A.S. has no personal pecuniary interests to disclose, other than being the lead of the German MS-Registry, which receives funding from a range of public and corporate sponsors, recently including: The German Innovation Fund (G-BA), The German MS Trust, The German Retirement Insurance, German MS Society, Biogen, Celgene (BMS), Merck, Novartis, Roche and Sanofi. P.D. is a full-time employee and a shareholder of Biogen. E.H.M.-L. and K.P. have no conflicts of interest to disclose. The views expressed in this article are the personal views of the author(s) and may not be understood or quoted as being made on behalf of or reflecting the position of the EMA or one of its committees or working parties. P.I. has served on scientific advisory boards for Biogen Idec, Bayer, Teva, Roche, Merck Serono, Novartis and Genzyme and has received funding for travel and/or Speaker honoraria from Sanofi Aventis, Genzyme, Biogen Idec, Teva, Merck Serono, Alexion and Novartis. J.I.R. has received honoraria from Novartis as a scientific advisor. He has received travel grants and attended courses and conferences on behalf of Merck-Serono Argentina, Novartis, Roche, Sanofi-Genzyme, Biogen, Bayer and Teva. He receives unrestricted research funding from Novartis, Biogen, Roche, Merck and Sanofi. M.M. has served in scientific advisory board for Sanofi, Novartis, Merck, and has received honoraria for lecturing from Biogen, Merck, Novartis, Roche, Genzyme, Bristol-Myers Squibb. She received research support and support for congress participation from Biogen, Genzyme, Roche, Merck, Novartis. A.v.d.W. has received honoraria and unrestricted research funding from Novartis, Biogen, Roche, Merck and Sanofi. G.C. reports that he has received consulting and speaking fees from Novartis, Teva Pharmaceutical Industries Ltd, Teva Italia Srl, Sanofi-Genzyme Corporation, Genzyme Europe, Merck KGgA, Merck Serono SpA, Celgene Group, Biogen Idec, Biogen Italia Srl, F. Hoffman-La Roche, Roche SpA, Almirall SpA, Forward Pharma, Medday, and Excemed. L.M.P. has no conflict to report related to this work other than being the chair and the coordinator of the MSDA initiative, receiving funding from a range of corporate sponsors, including Biogen, Bristol-Myers Squibb, Janssen Pharmaceuticals, Merck, Novartis, and Roche. All other co-authors have no relevant competing interests to report. The authors declare that the funding sources did not influence the content of this work.

Multiple Sclerosis (MS) is an inflammatory autoimmune disease of the central nervous system, causing increased vulnerability to infections and disability among young adults. Ever since the outbreak of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 infections, there have been concerns among people with MS (PwMS) about the potential interactions between various disease-modifying therapies and COVID-19. The COVID-19 in MS Global Data Sharing Initiative (GDSI) was initiated in 2020 with the aim of addressing these concerns. This paper focuses on the anonymisation and publicly releasing of a GDSI sub-dataset, comprising data entered by PwMS and clinicians using a fast data entry tool. The dataset includes information on demographics, comorbidities and hospital stay and COVID-19 symptoms of PwMS. The dataset can be used to perform different statistical analyses to improve our understanding of COVID-19 in MS. Furthermore, this dataset can also be used within the context of educational activities to educate different stakeholders on the complex data science topics that were used within the GDSI., (© 2024. The Author(s).)

Background: Investigating low-prevalence diseases such as multiple sclerosis is challenging because of the rather small number of individuals affected by this disease and the scattering of real-world data across numerous data sources. These obstacles impair data integration, standardization, and analysis, which negatively impact the generation of significant meaningful clinical evidence., Objective: This study aims to present a comprehensive, research question-agnostic, multistakeholder-driven end-to-end data analysis pipeline that accommodates 3 prevalent data-sharing streams: individual data sharing, core data set sharing, and federated model sharing., Methods: A demand-driven methodology is employed for standardization, followed by 3 streams of data acquisition, a data quality enhancement process, a data integration procedure, and a concluding analysis stage to fulfill real-world data-sharing requirements. This pipeline's effectiveness was demonstrated through its successful implementation in the COVID-19 and multiple sclerosis global data sharing initiative., Results: The global data sharing initiative yielded multiple scientific publications and provided extensive worldwide guidance for the community with multiple sclerosis. The pipeline facilitated gathering pertinent data from various sources, accommodating distinct sharing streams and assimilating them into a unified data set for subsequent statistical analysis or secure data examination. This pipeline contributed to the assembly of the largest data set of people with multiple sclerosis infected with COVID-19., Conclusions: The proposed data analysis pipeline exemplifies the potential of global stakeholder collaboration and underlines the significance of evidence-based decision-making. It serves as a paradigm for how data sharing initiatives can propel advancements in health care, emphasizing its adaptability and capacity to address diverse research inquiries., (©Ashkan Pirmani, Edward De Brouwer, Lotte Geys, Tina Parciak, Yves Moreau, Liesbet M Peeters. Originally published in JMIR Medical Informatics (https://medinform.jmir.org), 09.11.2023.)

Background: Cladribine tablets are a highly effective immune reconstitution therapy licensed for treating relapsing multiple sclerosis (RMS) in Europe since 2017. Currently, there is a high demand for real-world data from different clinical settings on the effectiveness and safety profile of cladribine in MS., Methods: Within this report, we retrospectively evaluated the outcomes of RMS patients who received cladribine between August 2018 and November 2021 at our Belgian institute. Patients with data for three effectiveness endpoints, more specifically, relapses, MRI observations, and confirmed disability worsening were incorporated into the analysis of 'no evidence of disease activity' (NEDA-3) re-baselined at 3 months. Safety endpoints included lymphopenia, liver transaminases, and adverse events (AEs) during follow-up. Descriptive statistics and time-to-event analysis were performed, including subgroup analysis by pre-treatment., Results: Of the 84 RMS patients included in this study (age 42 [33-50], 64.3% female, diagnosis duration 6 [2-11] years, baseline EDSS 2.5 [1.5-3.6]), 14 (16.7%) patients experienced relapses, while disability progression and brain MRI activity occurred in 8.5% (6/71) and 6.3% (5/79). This resulted in 72.6% (n = 69, standard error 6%) retaining NEDA-3 status at the mean follow-up time of 22.6 ± 11.5 months. During the first year after cladribine initiation, disease activity prevailed more in patients with ≥2 prior DMTs and those switching from fingolimod, although both trends were not statistically significant. In terms of safety, 67.9% reported at least one AE during follow-up, the most frequent being fatigue (64.9%) and skin-related problems (38.6%)., Conclusion: Overall, our research results confirm cladribine's safety and effectiveness among RMS patients in real-world conditions. After the re-baseline, we observed high rates of NEDA-3-retention, and no new safety signals were noted., Competing Interests: Declaration of Competing Interest Bart Van Wijmeersch has received honoraria, travel, and research grants from Almirall, BMS, Biogen, Imcyse, Janssen, Merck Serono, Novartis, Roche, Sanofi Genzyme, and Teva Pharmaceuticals. Veronica Popescu has received honoraria, travel, and research grants from Almirall, Biogen, Medtronic, Merck, Novartis, Roche, Sanofi Genzyme, and Teva Pharmaceuticals. The remaining authors have no disclosures related to this study., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Task-free functional connectivity in animal models provides an experimental framework to examine connectivity phenomena under controlled conditions and allows for comparisons with data modalities collected under invasive or terminal procedures. Currently, animal acquisitions are performed with varying protocols and analyses that hamper result comparison and integration. Here we introduce StandardRat, a consensus rat functional magnetic resonance imaging acquisition protocol tested across 20 centers. To develop this protocol with optimized acquisition and processing parameters, we initially aggregated 65 functional imaging datasets acquired from rats across 46 centers. We developed a reproducible pipeline for analyzing rat data acquired with diverse protocols and determined experimental and processing parameters associated with the robust detection of functional connectivity across centers. We show that the standardized protocol enhances biologically plausible functional connectivity patterns relative to previous acquisitions. The protocol and processing pipeline described here is openly shared with the neuroimaging community to promote interoperability and cooperation toward tackling the most important challenges in neuroscience., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Background and Objectives: Certain demographic and clinical characteristics, including the use of some disease-modifying therapies (DMTs), are associated with severe acute respiratory syndrome coronavirus 2 infection severity in people with multiple sclerosis (MS). Comprehensive exploration of these relationships in large international samples is needed., Methods: Clinician-reported demographic/clinical data from 27 countries were aggregated into a data set of 5,648 patients with suspected/confirmed coronavirus disease 2019 (COVID-19). COVID-19 severity outcomes (hospitalization, admission to intensive care unit [ICU], requiring artificial ventilation, and death) were assessed using multilevel mixed-effects ordered probit and logistic regression, adjusted for age, sex, disability, and MS phenotype. DMTs were individually compared with glatiramer acetate, and anti-CD20 DMTs with pooled other DMTs and with natalizumab., Results: Of 5,648 patients, 922 (16.6%) with suspected and 4,646 (83.4%) with confirmed COVID-19 were included. Male sex, older age, progressive MS, and higher disability were associated with more severe COVID-19. Compared with glatiramer acetate, ocrelizumab and rituximab were associated with higher probabilities of hospitalization (4% [95% CI 1-7] and 7% [95% CI 4-11]), ICU/artificial ventilation (2% [95% CI 0-4] and 4% [95% CI 2-6]), and death (1% [95% CI 0-2] and 2% [95% CI 1-4]) (predicted marginal effects). Untreated patients had 5% (95% CI 2-8), 3% (95% CI 1-5), and 1% (95% CI 0-3) higher probabilities of the 3 respective levels of COVID-19 severity than glatiramer acetate. Compared with pooled other DMTs and with natalizumab, the associations of ocrelizumab and rituximab with COVID-19 severity were also more pronounced. All associations persisted/enhanced on restriction to confirmed COVID-19., Discussion: Analyzing the largest international real-world data set of people with MS with suspected/confirmed COVID-19 confirms that the use of anti-CD20 medication (both ocrelizumab and rituximab), as well as male sex, older age, progressive MS, and higher disability are associated with more severe course of COVID-19., (Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology.)

Multiple sclerosis (MS) is a chronic disease affecting millions of people worldwide. Through the demyelinating and axonal pathology of MS, the signal conduction in the central nervous system is affected. Evoked potential measurements allow clinicians to monitor this process and can be used for decision support. We share a dataset that contains motor evoked potential (MEP) measurements, in which the brain is stimulated and the resulting signal is measured in the hands and feet. This results in time series of 100 milliseconds long. Typically, both hands and feet are measured in one hospital visit. The dataset contains 5586 visits of 963 patients, performed in day-to-day clinical care over a period of 6 years. The dataset consists of approximately 100,000 MEP. Clinical metadata such as the expanded disability status scale, sex, and age is also available. This dataset can be used to explore the role of evoked potentials in MS research and patient care. It may also be used as a benchmark for time series analysis and predictive modelling., (© 2022. The Author(s).)

Background: One of the major objectives of the Multiple Sclerosis Data Alliance (MSDA) is to enable better discovery of multiple sclerosis (MS) real-world data (RWD)., Methods: We implemented the MSDA Catalogue, which is available worldwide. The current version of the MSDA Catalogue collects descriptive information on governance, purpose, inclusion criteria, procedures for data quality control, and how and which data are collected, including the use of e-health technologies and data on collection of COVID-19 variables. The current cataloguing procedure is performed in several manual steps, securing an effective catalogue., Results: Herein we summarize the status of the MSDA Catalogue as of January 6, 2021. To date, 38 data sources across five continents are included in the MSDA Catalogue. These data sources differ in purpose, maturity, and variables collected, but this landscaping effort shows that there is substantial alignment on some domains. The MSDA Catalogue shows that personal data and basic disease data are the most collected categories of variables, whereas data on fatigue measurements and cognition scales are the least collected in MS registries/cohorts., Conclusions: The Web-based MSDA Catalogue provides strategic overview and allows authorized end users to browse metadata profiles of data cohorts and data sources. There are many existing and arising RWD sources in MS. Detailed cataloguing of MS RWD is a first and useful step toward reducing the time needed to discover MS RWD sets and promoting collaboration., Competing Interests: Financial Disclosures: Dr Geys is funded by the Flemish Government under the “Onderzoeksprogramma Artificiële Intelligentie Vlaanderen.” Ms Parciak and Mr Pirmani are funded by the Fonds voor Wetenschappelijk Onderzoek (FWO) (project R4859). Dr McBurney is the chief research officer and Ms Schmidt is the vice president of scientific operations of the Accelerated Cure Project for MS (ACP). Dr McBurney has received consulting payments from EMD Serono, which have been donated to the ACP. The ACP has received grants, collaboration funding, payments for use of assets, or in-kind contributions from the following companies: EMD Serono, Novartis, Sanofi/Genzyme, Biogen, Genentech, AbbVie, Octave, GlycoMinds, Pfizer, MedDay, AstraZeneca, Teva, Mallinckrodt, MSDx, Regeneron Genetics Center, BC Platforms, and Bristol Myers Squibb (Celgene). The ACP has also received funding from the Patient-Centered Outcomes Research Institute, the National MS Society, and the Italian MS Society. Dr Rojas has received reimbursement for developing educational presentations, educational and research grants, consultation fees, and travel stipends from Biogen, Genzyme, Merck, and Novartis. Dr García-Merino has received honoraria for travel expenses, lecturing, or consulting from Merck, Biogen, Bayer, Teva, Roche, Novartis, Genzyme, and Almirall and research grants from Biogen and Novartis. Dr Fernández has received travel support and/or research grants and/or lecture fees and/or advisory services from Merck, Bayer, Biogen, Teva, Sanofi Genzyme, Novartis, Roche, Almirall, Allergan, Orizon, and Araclon. Dr Gobbi’s employer (Department of Neurology, Regional Hospital Lugano [EOC], Switzerland) receives financial support for speaking or educational, research, or travel grants from AbbVie, Almirall, Biogen, Celgene, Sanofi, Merck, Novartis, Teva, and Roche. The Neuroepidemiology Unit and HOLISM-related work (Drs Simpson-Yap and Nag) was supported by anonymous philanthropic funders and Mr Wal Pisciotta. Dr Yamout has received speaker honoraria from Bayer, Biogen, Genesis Pharma, Genpharm, Genzyme, Merck, and Novartis; research grants from Bayer, Biogen, Merck, Novartis, and Pfizer; and advisory board honoraria from Bayer, Biogen, Genzyme, Merck, Novartis, and Genpharm. The Swiss MS Registry (Dr Steinemann) is funded by a grant from the Swiss Multiple Sclerosis Society. Dr Seeldrayers has been responsible for BELTRIMS in the frame of the Belgian Study Group for Multiple Sclerosis; in the frame of her clinical activities, she has received travel grants and honoraria for lectures and participation in advisory boards from Biogen, Bayer, Merck, Teva, Sanofi Genzyme, Novartis, and Roche. Dr Dubois is a clinical investigator of the Research Fund Flanders (FWO-Vlaanderen). Mr Stahmann is the lead of the German MS Registry, which receives funding from a range of public and corporate sponsors, recently including the German Innovation Fund, German MS Trust, German MS Society, Biogen, Celgene (Bristol Myers Squibb), Merck, Novartis, Roche, and Sanofi. Dr Drulovic has received travel support and/or research grants and/or lecture fees and/or advisory services from Novartis, Bayer, Merck, Sanofi Genzyme, Roche, Teva, Medis, Hemofarm, and Medtronic. Dr Pekmezovic has received travel support and/or research grants and/or lecture fees and/or advisory services from Novartis, Merck, Sanofi Genzyme, Roche, Teva, and Hemofarm. Dr Tintore discloses consulting services and speaking honoraria from Almirall, Bayer Schering Pharma, Biogen, Genzyme, Janssen, Merck-Serono, Novartis, Roche, Sanofi-Aventis, Viela Bio, and Teva and research grants and support from Carlos III Health Institute, Foundation Genzyme, Foundation Salud 2000, Biogen, and Novartis. Dr Kalkers received speaking fees and consulting fees from Novartis, Teva, and Sanofi Genzyme. The Dutch MS Registry is funded by a project from ZonMW and co-financed by Biogen, Celgene, Merck, Mylan, Novartis, Roche, Sanofi Genzyme, and Teva. Dr Zakaria has received fees for lectures and advisory board meetings from Merck, Roche, Sanofi, Biogen, Novartis, and Bayer. Dr Naseer received speaker honoraria and served on scientific advisory boards for Novartis, Roche, Merck, Sanofi Genzyme, and Bayer. Dr Grigoriadis has received travel support and/or honoraria and/or research grants and/or lecture fees and/or advisory services and/or consulting fees from Novartis, Bayer, Merck, Genesis, Sanofi, Specifar, Roche, Biogen, Teva, Mylan, Merck Serono, Sanofi Genzyme, Celgene, and ELPEN. Dr Boziki has received travel support and/or research grants and/or lecture fees and/or advisory services from the Hellenic Foundation for Research and Innovation, the Ministry of Education’s Education and Lifelong Learning Program, the Hellenic Neurological Society, the Hellenic Academy of Neuroimmunology, Biogen, Novartis, Teva, Bayer, Genesis Pharma, Sanofi, Specifar, Roche, and Merck. Dr Pawlak has received grants from the Polish National Science Centre during the conduct of the study as well as personal fees from Roche, Novartis, Merck, and Biogen outside the submitted work. Dr Dobson has received travel support and/or research grants and/or consulting fees from Biogen, Roche, Celgene, Sanofi Genzyme, Teva, and Merck. Dr Hellwig has received travel support and/or research grants and/or lecture fees and/or advisory services from Novartis, Bayer, Merck, Sanofi Genzyme, Roche, Biogen, Teva, and Mylan. Dr Leocani has received consulting fees and/or research support from Biogen, Merck KGgA, Novartis, Hoffman-La Roche, Almirall SpA, and Bristol Myers Squibb. Dr Van Wijmeersch has received travel support and/or research grants and/or lecture fees and/or advisory services from Almirall, Biogen, Bristol Myers Squibb, Janssen Pharmaceutica, Merck, Novartis, Roche, Sanofi, and Teva. Dr Peeters is chair of the MSDA, which receives income from a range of corporate sponsors, including Biogen, Bristol Myers Squibb, Janssen, Merck KGaA, Mylan, Novartis, QMENTA, and Roche and has received funding from the Flemish Government under the “Onderzoeksprogramma Artificiële Intelligentie Vlaanderen.” The other authors declare no conflicts of interest., (© 2021 Consortium of Multiple Sclerosis Centers.)

Background: The COVID-19 pandemic has resulted in uncertain access to medical treatment for people with multiple sclerosis (pwMS) all over the world. However, there is no data regarding its impact on access to health care of pwMS from Latin America., Objectives: We investigated and described changes in health care delivery for pwMS from Latin America during the COVID-19 pandemic., Methods: PwMS from 18 patient organizations of the region completed a web-based survey hosted from May to October 2020., Results: A total of 602 pwMS completed the questionnaire. Changes in disease-modifying therapies (DMTs) use: 6.7% of pwMS on continuous DMTs claimed to stopped them; 14.1% of those on infusion therapies declared to postpone their dosing; 68.8% declared delaying the initiation of a DMT. Disruptions in accessing rehabilitation services were reported by 65.7%. Changes in laboratory and MRI monitoring were reported by 30% and 33%, respectively. In a multivariable-adjusted logistic regression model, changes in laboratory monitoring were significantly associated with increased odds of postponing MRI monitoring (OR 4.09 CI95% 2.79-6.00, p < 0.001)., Conclusions: The COVID-19 pandemic has disrupted all aspects of the routine care for pwMS from Latin America. Consequences are yet to be determined., (Copyright © 2021. Published by Elsevier B.V.)

Background: There is no data regarding COVID-19 in Multiple Sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) patients in Latin America., Objective: The objective of this study was to describe the clinical characteristics and outcomes of patients included in RELACOEM, a LATAM registry of MS and NMOSD patients infected with COVID-19., Methods: RELACOEM is a longitudinal, strictly observational registry of MS and NMOSD patients who suffer COVID-19 and Dengue in LATAM. Inclusion criteria to the registry were either: (1) a biologically confirmed COVID-19 diagnosis based on a positive result of a COVID-19 polymerase chain reaction (PCR) test on a nasopharyngeal swab; or (2) COVID-19-typical symptoms (triad of cough, fever, and asthenia) in an epidemic zone of COVID-19. Descriptive statistics were performed on demographic and clinical variables. The cohort was later stratified for MS and NMOSD and univariate and multivariate logistic regression analysis was performed to identify variables associated with hospitalizations/intensive critical units (ICU) admission., Results: 145 patients were included in the registry from 15 countries and 51 treating physicians. A total of 129 (89%) were MS patients and 16 (11%) NMOSD. 81.4% patients had confirmed COVID-19 and 18.6% were suspected cases. 23 (15.8%) patients were hospitalized, 9 (6.2%) required ICU and 5 (3.4 %) died due to COVID-19. In MS patients, greater age (OR 1.17, 95% CI 1.05 - 1.25) and disease duration (OR 1.39, 95%CI 1.14-1.69) were associated with hospitalization/ICU. In NMOSD patients, a greater age (54.3 vs. 36 years, p=<0.001), increased EDSS (5.5 vs 2.9, p=0.0012) and disease duration (18.5 vs. 10.3 years, p=0.001) were significantly associated with hospitalization/ICU., Conclusion: we found that in MS patients, age and disease duration was associated with hospitalization and ICU admission requirement, while age, disease duration and EDSS was associated in NMOSD., Competing Interests: Disclosure of conflicts of interest Edgardo Cristiano has received fees for consultations as a scientific advisory board member and for travel to meetings, conferences, and clinical trials of the following companies: Avanir, Bayer, Biogen, Merck, Novartis, Roche and Teva. Juan Ignacio Rojas has received honoraria from Novartis as a scientific advisor. He has received travel grants and attended courses and conferences on behalf of Merck-Serono Argentina, Novartis Argentina. Liliana Patrucco has received honoraria for scientific and research grants from Teva Tuteur, Merck Serono, Biogen Idec, and Bayer Schering. Ricardo Alonso and Berenice Silva has received personal compensation for consulting, serving on a scientific advisory board, speaking, or other activities from Biogen, Merck Serono, Novartis, Sanofi -Genzyme and Roche. Orlando Garcea has received personal compensation for consulting, serving on a scientific advisory board, speaking, or other activities from Biogen, Merck Serono, Novartis, Sanofi -Genzyme and Roche. Liesbet M. Peeters has no personal pecuniary interests to disclose, other than being the chair of The MS Data Alliance (MSDA), which receives income from a range of corporate sponsors, recently including: Biogen, BristolMyersSquibb (formerly Celgene), Canopy Growth Corporation, Genzyme, Icometrix, Merck, Mylan, Novartis, QMENTA, Quanterix, Roche. The rest of authors declares no conflict of interest with the study project., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Cytotoxic CD4 + T cells (CD4 CTL) are terminally differentiated T helper cells that contribute to autoimmune diseases, such as multiple sclerosis. We developed a novel triple co-culture transwell assay to study mutual interactions between CD4 CTL, conventional TH cells, and regulatory T cells (Tregs) simultaneously. We show that, while CD4 CTL are resistant to suppression by Tregs in vitro, the conditioned medium of CD4 CTL accentuates the suppressive phenotype of Tregs by upregulating IL-10, Granzyme B, CTLA-4, and PD-1. We demonstrate that CD4 CTL conditioned medium skews memory TH cells to a TH17 phenotype, suggesting that the CD4 CTL induce bystander polarization. In our triple co-culture assay, the CD4 CTL secretome promotes the proliferation of TH cells, even in the presence of Tregs. However, when cell-cell contact is established between CD4 CTL and TH cells, the proliferation of TH cells is no longer increased and Treg-mediated suppression is restored. Taken together, our results suggest that when TH cells acquire cytotoxic properties, these Treg-resistant CD4 CTL affect the proliferation and phenotype of conventional TH cells in their vicinity. By creating such a pro-inflammatory microenvironment, CD4 CTL may favor their own persistence and expansion, and that of other potentially pathogenic TH cells, thereby contributing to pathogenic responses in autoimmune disorders.

The anterior cingulate area (ACC) is an integral part of the prefrontal cortex in mice and supports cognitive functions, including attentional processes, motion planning and execution as well as remote memory, fear and pain. Previous anatomical and functional imaging studies demonstrated that the ACC is interconnected with numerous brain regions, such as motor and sensory cortices, amygdala and limbic areas, suggesting it serves as a hub in functional networks. However, the exact role of the ACC in regulating functional network activity and connectivity remains to be elucidated. Recently developed neuromodulatory techniques, such as Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) allow for precise control of neuronal activity. In this study, we used an inhibitory kappa-opioid receptor DREADD (KORD) to temporally inhibit neuronal firing in the right ACC of mice and assessed functional network activity and connectivity using non-invasive functional magnetic resonance imaging (MRI). We demonstrated that KORD-induced inhibition of the right ACC induced blood oxygenation-level dependent (BOLD) signal decreases and increases in connected brain regions of both hemispheres. More specifically, altered neuronal activity could be observed in functional brain networks including connections with sensory cortex, thalamus, basolateral amygdala and ventral pallidum, areas involved in attention processes, working memory, fear behavior and reward respectively. Furthermore, these modulations in neuronal activity were associated with decreased intra- and interhemispheric functional connectivity. Our results consolidate the hub role of the mouse ACC in functional networks and further demonstrate that the combination of the DREADD technology and non-invasive functional imaging methods is a valuable tool for unraveling mechanisms of network function and dysfunction by reversible inactivation of selected targets., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: We need high-quality data to assess the determinants for COVID-19 severity in people with MS (PwMS). Several studies have recently emerged but there is great benefit in aligning data collection efforts at a global scale., Objectives: Our mission is to scale-up COVID-19 data collection efforts and provide the MS community with data-driven insights as soon as possible., Methods: Numerous stakeholders were brought together. Small dedicated interdisciplinary task forces were created to speed-up the formulation of the study design and work plan. First step was to agree upon a COVID-19 MS core data set. Second, we worked on providing a user-friendly and rapid pipeline to share COVID-19 data at a global scale., Results: The COVID-19 MS core data set was agreed within 48 hours. To date, 23 data collection partners are involved and the first data imports have been performed successfully. Data processing and analysis is an on-going process., Conclusions: We reached a consensus on a core data set and established data sharing processes with multiple partners to address an urgent need for information to guide clinical practice. First results show that partners are motivated to share data to attain the ultimate joint goal: better understand the effect of COVID-19 in PwMS.

The discovery of the default mode network (DMN), a large-scale brain network that is suppressed during attention-demanding tasks, had major impact in neuroscience. This network exhibits an antagonistic relationship with attention-related networks. A better understanding of the processes underlying modulation of DMN is imperative, as this network is compromised in several neurological diseases. Cholinergic neuromodulation is one of the major regulatory networks for attention, and studies suggest a role in regulation of the DMN. In this study, we unilaterally activated the right basal forebrain cholinergic neurons and observed decreased right intra-hemispheric and interhemispheric FC in the default mode like network (DMLN). Our findings provide critical insights into the interplay between cholinergic neuromodulation and DMLN, demonstrate that differential effects can be exerted between the two hemispheres by unilateral stimulation, and open windows for further studies involving directed modulations of DMN in treatments for diseases demonstrating compromised DMN activity., Competing Interests: The authors declare no competing interests., (© 2020 The Author(s).)

Motor Evoked Potentials (MEPs) are used to monitor disability progression in multiple sclerosis (MS). Their morphology plays an important role in this process. Currently, however, there is no clear definition of what constitutes a normal or abnormal morphology. To address this, five experts independently labeled the morphology (normal or abnormal) of the same set of 1,000 MEPs. The intra- and inter-rater agreement between the experts indicates they agree on the concept of morphology, but differ in their choice of threshold between normal and abnormal morphology. We subsequently performed an automated extraction of 5,943 time series features from the MEPs to identify a valid proxy for morphology, based on the provided labels. To do this, we compared the cross-validation performances of one-dimensional logistic regression models fitted to each of the features individually. We find that the approximate entropy (ApEn) feature can accurately reproduce the majority-vote labels. The performance of this feature is evaluated on an independent test set by comparing to the majority vote of the neurologists, obtaining an AUC score of 0.92. The model slightly outperforms the average neurologist at reproducing the neurologists consensus-vote labels. We can conclude that MEP morphology can be consistently defined by pooling the interpretations from multiple neurologists and that ApEn is a valid continuous score for this. Having an objective and reproducible MEP morphological abnormality score will allow researchers to include this feature in their models, without manual annotation becoming a bottleneck. This is crucial for large-scale, multi-center datasets. An exploratory analysis on a large single-center dataset shows that ApEn is potentially clinically useful. Introducing an automated, objective, and reproducible definition of morphology could help overcome some of the barriers that are currently obstructing broad adoption of evoked potentials in daily care and patient follow-up, such as standardization of measurements between different centers, and formulating guidelines for clinical use., (Copyright © 2020 Yperman, Becker, Valkenborg, Hellings, Cambron, Dive, Laureys, Popescu, Van Wijmeersch and Peeters.)

Background: Evoked potentials (EPs) are a measure of the conductivity of the central nervous system. They are used to monitor disease progression of multiple sclerosis patients. Previous studies only extracted a few variables from the EPs, which are often further condensed into a single variable: the EP score. We perform a machine learning analysis of motor EP that uses the whole time series, instead of a few variables, to predict disability progression after two years. Obtaining realistic performance estimates of this task has been difficult because of small data set sizes. We recently extracted a dataset of EPs from the Rehabiliation & MS Center in Overpelt, Belgium. Our data set is large enough to obtain, for the first time, a performance estimate on an independent test set containing different patients., Methods: We extracted a large number of time series features from the motor EPs with the highly comparative time series analysis software package. Mutual information with the target and the Boruta method are used to find features which contain information not included in the features studied in the literature. We use random forests (RF) and logistic regression (LR) classifiers to predict disability progression after two years. Statistical significance of the performance increase when adding extra features is checked., Results: Including extra time series features in motor EPs leads to a statistically significant improvement compared to using only the known features, although the effect is limited in magnitude (ΔAUC = 0.02 for RF and ΔAUC = 0.05 for LR). RF with extra time series features obtains the best performance (AUC = 0.75±0.07 (mean and standard deviation)), which is good considering the limited number of biomarkers in the model. RF (a nonlinear classifier) outperforms LR (a linear classifier)., Conclusions: Using machine learning methods on EPs shows promising predictive performance. Using additional EP time series features beyond those already in use leads to a modest increase in performance. Larger datasets, preferably multi-center, are needed for further research. Given a large enough dataset, these models may be used to support clinicians in their decision making process regarding future treatment.

The combination of chemogenetics targeting specific brain cell populations with in vivo imaging techniques provides scientists with a powerful new tool to study functional neural networks at the whole-brain scale. A number of recent studies indicate the potential of this approach to increase our understanding of brain function in health and disease. In this review, we discuss the employment of a specific chemogenetic tool, designer receptors exclusively activated by designer drugs, in conjunction with non-invasive neuroimaging techniques such as PET and MRI. We highlight the utility of using this multiscale approach in longitudinal studies and its ability to identify novel brain circuits relevant to behaviour that can be monitored in parallel. In addition, some identified shortcomings in this technique and more recent efforts to overcome them are also presented. Finally, we discuss the translational potential of designer receptors exclusively activated by designer drugs in neuroimaging and the promise it holds for future neurotheranostic applications., (© 2019 The British Pharmacological Society.)

The default mode network is a large-scale brain network that is active during rest and internally focused states and deactivates as well as desynchronizes during externally oriented (top-down) attention demanding cognitive tasks. However, it is not sufficiently understood if salient stimuli, able to trigger bottom-up attentional processes, could also result in similar reduction of activity and functional connectivity in the DMN. In this study, we investigated whether bottom-up sensory processing could influence the default mode-like network (DMLN) in rats. DMLN activity was examined using block-design visual functional magnetic resonance imaging (fMRI) while its synchronization was investigated by comparing functional connectivity during a resting versus a continuously stimulated brain state by unpredicted light flashes. We demonstrated that the BOLD response in DMLN regions was decreased during visual stimulus blocks and increased during blanks. Furthermore, decreased inter-network functional connectivity between the DMLN and visual networks as well as decreased intra-network functional connectivity within the DMLN was observed during the continuous visual stimulation. These results suggest that triggering of bottom-up attention mechanisms in sedated rats can lead to a cascade similar to top-down orienting of attention in humans and is able to deactivate and desynchronize the DMLN., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Personalized treatment is highly desirable in multiple sclerosis (MS). We believe that multidisciplinary measurements including clinical, functional and patient-reported outcome measures in combination with extensive patient profiling can enhance personalized treatment and rehabilitation strategies. We elaborate on four reasons behind this statement: (1) MS disease activity and progression are complex and multidimensional concepts in nature and thereby defy a one-size-fits-all description, (2) functioning, progression, treatment, and rehabilitation effects are interdependent and should be investigated together, (3) personalized healthcare is based on the dynamics of system biology and on technology that confirms a patient's fundamental biology and (4) inclusion of patient-reported outcome measures can facilitate patient-relevant healthcare. We discuss currently available multidisciplinary MS data initiatives and introduce joint actions to further increase the overall success. With this topical review, we hope to drive the MS community to invest in expanding towards more multidisciplinary and longitudinal data collection.

Multiple sclerosis (MS) is a progressive demyelinating and degenerative disease of the central nervous system with symptoms depending on the disease type and the site of lesions and is featured by heterogeneity of clinical expressions and responses to treatment strategies. An individualized clinical follow-up and multidisciplinary treatment is required. Transforming the population-based management of today into an individualized, personalized and precision-level management is a major goal in research. Indeed, a complex and unique interplay between genetic background and environmental exposure in each case likely determines clinical heterogeneity. To reach insights at the individual level, extensive amount of data are required. Many databases have been developed over the last few decades, but access to them is limited, and data are acquired in different ways and differences in definitions and indexing and software platforms preclude direct integration. Most existing (inter)national registers and IT platforms are strictly observational or focus on disease epidemiology or access to new disease modifying drugs. Here, a method to revolutionize management of MS to a personalized, individualized and precision level is outlined. The key to achieve this next level is FAIR data.

Multiple sclerosis (MS) is the leading cause of chronic neurological disability in young adults. The clinical disease course of MS varies greatly between individuals, with some patients progressing much more rapidly than others, making prognosis almost impossible. We previously discovered that cytotoxic CD4+ T cells (CD4+ CTL), identified by the loss of CD28, are able to migrate to sites of inflammation and that they contribute to tissue damage. Furthermore, in an animal model for MS, we showed that these cells are correlated with inflammation, demyelination, and disability. Therefore, we hypothesize that CD4+ CTL drive progression of MS and have prognostic value. To support this hypothesis, we investigated whether CD4+ CTL are correlated with worse clinical outcome and evaluated the prognostic value of these cells in MS. To this end, the percentage of CD4+CD28null T cells was measured in the blood of 176 patients with relapsing-remitting MS (=baseline). Multimodal evoked potentials (EP) combining information on motoric, visual, and somatosensoric EP, as well as Kurtzke expanded disability status scale (EDSS) were used as outcome measurements at baseline and after 3 and 5 years. The baseline CD4+CD28null T cell percentage is associated with EP ( P  = 0.003, R 2  = 0.28), indicating a link between these cells and disease severity. In addition, the baseline CD4+CD28null T cell percentage has a prognostic value since it is associated with EP after 3 years ( P  = 0.005, R 2  = 0.29) and with EP and EDSS after 5 years ( P  = 0.008, R 2  = 0.42 and P  = 0.003, R 2  = 0.27). To the best of our knowledge, this study provides the first direct link between the presence of CD4+ CTL and MS disease severity, as well as its prognostic value. Therefore, we further elaborate on two important research perspectives: 1° investigating strategies to block or reverse pathways in the formation of these cells resulting in new treatments that slow down MS disease progression, 2° including immunophenotyping in prediction modeling studies to aim for personalized medicine.

Cytomegalovirus (CMV) is a latent virus which causes chronic activation of the immune system. Here, we demonstrate that cytotoxic and pro-inflammatory CD4 + CD28 null T cells are only present in CMV seropositive donors and that CMV-specific Immunoglobulin (Ig) G titers correlate with the percentage of these cells. In vitro stimulation of peripheral blood mononuclear cells with CMVpp65 peptide resulted in the expansion of pre-existing CD4 + CD28 null T cells. In vivo, we observed de novo formation, as well as expansion of CD4 + CD28 null T cells in two different chronic inflammation models, namely the murine CMV (MCMV) model and the experimental autoimmune encephalomyelitis (EAE) model for multiple sclerosis (MS). In EAE, the percentage of peripheral CD4 + CD28 null T cells correlated with disease severity. Pre-exposure to MCMV further aggravated EAE symptoms, which was paralleled by peripheral expansion of CD4 + CD28 null T cells, increased splenocyte MOG reactivity and higher levels of spinal cord demyelination. Cytotoxic CD4 + T cells were identified in demyelinated spinal cord regions, suggesting that peripherally expanded CD4 + CD28 null T cells migrate towards the central nervous system to inflict damage. Taken together, we demonstrate that CMV drives the expansion of CD4 + CD28 null T cells, thereby boosting the activation of disease-specific CD4 + T cells and aggravating autoimmune mediated inflammation and demyelination.

Genetic factors involved in susceptibility to insect bite hypersensitivity (IBH) in Belgian Warmblood horses (BWP) were investigated. Data relating to 3409 horses were collected using a questionnaire, administered to owners during sport competitions, BWP breeding days, breeder visits and after phone calls. Horses were classified as IBH-affected or unaffected, based on two 'disease classifiers': a lifetime record, based on owner information (life&#95;status) and another based on whether or not the horse was showing clinical signs at the time of questioning (clin&#95;status). IBH prevalence was 10% based on life&#95;status, and 6.2% based on clin&#95;status. The heritabilities estimated using threshold animal models varied from 0.65 to 0.78 on the underlying scale (0.18-0.26 on the observed scale). These research findings indicate that susceptibility to IBH is a heritable trait in BWP., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Genetic parameters for chronic progressive lymphedema (CPL)-associated traits in Belgian Draught Horses were estimated, using a multitrait animal model. Clinical scores of CPL in the four limbs/horse (CPLclin ), skinfold thickness and hair samples (hair diameter) were studied. Due to CPLclin uncertainty in younger horses (progressive CPL character), a restricted data set (D&#95;3+) was formed, excluding records from horses under 3 years from the complete data set (D&#95;full). Age, gender, coat colour and limb hair pigmentation were included as fixed, permanent environment and date of recording as random effects. Higher CPLclin certainty (D&#95;3+) increased heritability coefficients of, and genetic correlations between traits, with CPLclin heritabilities (SE) for the respective data sets: 0.11 (0.06) and 0.26 (0.05). A large proportion of the CPLclin variance was attributed to the permanent environmental effect in D&#95;full, but less in D&#95;3+. Date of recording explained a proportion of variance from 0.09 ± 0.03 to 0.61 ± 0.08. Additive genetic correlations between CPLclin and both skinfold thickness and hair diameter showed the latter two traits cannot be used as a direct diagnostic aid for CPL. Due to the relatively low heritability of CPLclin , selection should focus on estimated breeding values (from repeated clinical examinations) to reduce CPL occurrence in the Belgian Draught Horse., (© 2014 Blackwell Verlag GmbH.)

Insect bite hypersensitivity (IBH) in horses represents an immunoglobulin E (IgE)-mediated hypersensitivity to salivary antigens from biting midges (Culicoides spp.). The aim of this study was to evaluate and compare the performances of IgE ELISAs using recombinant Culicoides spp. Obsoletus group salivary gland antigens or crude whole body extracts ('ObsWBE'), C. nubeculosus recombinant proteins (Culn1, 3, 4, 5, 7, 8 and 10) and Obsoletus group recombinant proteins (Culo1 and 2). IgE levels were measured in plasma of 343 Warmblood horses classified as IBH-affected (n=167) and IBH-unaffected (n=176) according to the owners' descriptions. IBH-affected horses were subdivided based on the severity of their clinical signs at sampling and whether or not their IBH history was considered to be classical. The accuracies of the tests increased when clinical signs at sampling were more pronounced or when the IBH history could be considered as classical. A combination of IgE levels against the three best performing Culicoides spp. recombinant proteins (Culn4, Culo1 and Culo2) and ObsWBE resulted in the best performing test. When IBH-affected horses showing a classical history of the disease and severe clinical signs were compared with IBH-unaffected horses, the Youden's index at the optimal cut-off for the three tests in combination was 0.67. This optimal cut-off had a sensitivity of 70%, a specificity of 97% and a total accuracy of 92%. The performance of the IgE ELISA was affected by the severity of IBH clinical signs at sampling and was improved when IgE levels against several recombinant proteins were combined., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The lysis cassette of Pseudomonas aeruginosa phage ϕKMV encodes a holin, endolysin, Rz and Rz1 in the canonical order. It has a tight organization with a high degree of overlapping genes and is highly conserved (between 96 and 100% identity at the protein level) among several other members of the "phiKMV-like viruses." The endolysin KMV45 exhibits characteristics as expected for a signal-arrest-release (SAR) endolysin, whereas the holin KMV44 is a typical pinholin. KMV45 is initially secreted as an inactive, membrane-anchored endolysin, which is subsequently released by membrane depolarization driven by the pinholin KMV44. The SAR domain of KMV45 is necessary for its full enzymatic activity, suggesting a refolding of the catalytic cleft upon release from the membrane. The physical proximity of the catalytic glutamic acid residue close to SAR domain suggests an alternative activation mechanism compared to the SAR endolysin of phages P1, ERA103 and 21. Expression of KMV44 leads to a quick cell lysis when paired with SAR endolysin KMV45, but not with the cytoplasmic phage λ endolysin, indicating the membrane depolarizing function of KMV44 rather than the large hole-making function characteristic of classical holins.