Ratiometric array of conjugated polymers–fluorescent protein provides a robust mammalian cell sensor

© 2016 American Chemical Society.Supramolecular complexes of a family of positively charged conjugated polymers (CPs) and green fluorescent protein (GFP) create a fluorescence resonance energy transfer (FRET)-based ratiometric biosensor array. Selective multivalent interactions of the CPs with mammalian cell surfaces caused differential change in FRET signals, providing a fingerprint signature for each cell type. The resulting fluorescence signatures allowed the identification of 16 different cell types and discrimination between healthy, cancerous, and metastatic cells, with the same genetic background. While the CP-GFP sensor array completely differentiated between the cell types, only partial classification was achieved for the CPs alone, validating the effectiveness of the ratiometric sensor. The utility of the biosensor was further demonstrated in the detection of blinded unknown samples, where 121 of 128 samples were correctly identified. Notably, this selectivity-based sensor stratified diverse cell types in minutes, using only 2000 cells, without requiring specific biomarkers or cell labeling

Antibodies Elicited in Response to EBNA-1 May Cross-React with dsDNA

Several genetic and environmental factors have been linked to Systemic Lupus Erythematosus (SLE). One environmental trigger that has a strong association with SLE is the Epstein Barr Virus (EBV). Our laboratory previously demonstrated that BALB/c mice expressing the complete EBNA-1 protein can develop antibodies to double stranded DNA (dsDNA). The present study was undertaken to understand why anti-dsDNA antibodies arise during the immune response to EBNA-1.In this study, we demonstrated that mouse antibodies elicited in response to EBNA-1 cross-react with dsDNA. First, we showed that adsorption of sera reactive with EBNA-1 and dsDNA, on dsDNA cellulose columns, diminished reactivity with EBNA-1. Next, we generated monoclonal antibodies (MAbs) to EBNA-1 and showed, by several methods, that they also reacted with dsDNA. Examination of two cross-reactive MAbs--3D4, generated in this laboratory, and 0211, a commercial MAb--revealed that 3D4 recognizes the carboxyl region of EBNA-1, while 0211 recognizes both the amino and carboxyl regions. In addition, 0211 binds moderately well to the ribonucleoprotein, Sm, which has been reported by others to elicit a cross-reactive response with EBNA-1, while 3D4 binds only weakly to Sm. This suggests that the epitope in the carboxyl region may be more important for cross-reactivity with dsDNA while the epitope in the amino region may be more important for cross-reactivity with Sm.In conclusion, our results demonstrate that antibodies to the EBNA-1 protein cross-react with dsDNA. This study is significant because it demonstrates a direct link between the viral antigen and the development of anti-dsDNA antibodies, which are the hallmark of SLE. Furthermore, it illustrates the crucial need to identify the epitopes in EBNA-1 responsible for this cross-reactivity so that therapeutic strategies can be designed to mask these regions from the immune system following EBV exposure

Concepts of Cardiac Development in Retrospect

Recent research, enabled by powerful molecular techniques, has revolutionized our concepts of cardiac development. It was firmly established that the early heart tube gives rise to the left ventricle only, and that the remainder of the myocardium is recruited from surrounding mesoderm during subsequent development. Also, the cardiac chambers were shown not to be derived from the entire looping heart tube, but only from the myocardium at its outer curvatures. Intriguingly, many years ago, classic experimental embryological studies reached very similar conclusions. However, with the current scientific emphasis on molecular mechanisms, old morphological insights became underexposed. Since cardiac development occurs in an architecturally complex and dynamic fashion, molecular insights can only fully be exploited when placed in a proper morphological context. In this communication we present excerpts of important embryological studies of the pioneers of experimental cardiac embryology of the previous century, to relate insights from the past to current observations

Mapping age- and sex-specific HIV prevalence in adults in sub-Saharan Africa, 2000–2018

Background: Human immunodeficiency virus and acquired immune deficiency syndrome (HIV/AIDS) is still among the leading causes of disease burden and mortality in sub-Saharan Africa (SSA), and the world is not on track to meet targets set for ending the epidemic by the Joint United Nations Programme on HIV/AIDS (UNAIDS) and the United Nations Sustainable Development Goals (SDGs). Precise HIV burden information is critical for effective geographic and epidemiological targeting of prevention and treatment interventions. Age- and sex-specific HIV prevalence estimates are widely available at the national level, and region-wide local estimates were recently published for adults overall. We add further dimensionality to previous analyses by estimating HIV prevalence at local scales, stratified into sex-specific 5-year age groups for adults ages 15–59 years across SSA. Methods: We analyzed data from 91 seroprevalence surveys and sentinel surveillance among antenatal care clinic (ANC) attendees using model-based geostatistical methods to produce estimates of HIV prevalence across 43 countries in SSA, from years 2000 to 2018, at a 5 × 5-km resolution and presented among second administrative level (typically districts or counties) units. Results: We found substantial variation in HIV prevalence across localities, ages, and sexes that have been masked in earlier analyses. Within-country variation in prevalence in 2018 was a median 3.5 times greater across ages and sexes, compared to for all adults combined. We note large within-district prevalence differences between age groups: for men, 50% of districts displayed at least a 14-fold difference between age groups with the highest and lowest prevalence, and at least a 9-fold difference for women. Prevalence trends also varied over time; between 2000 and 2018, 70% of all districts saw a reduction in prevalence greater than five percentage points in at least one sex and age group. Meanwhile, over 30% of all districts saw at least a five percentage point prevalence increase in one or more sex and age group. Conclusions: As the HIV epidemic persists and evolves in SSA, geographic and demographic shifts in prevention and treatment efforts are necessary. These estimates offer epidemiologically informative detail to better guide more targeted interventions, vital for combating HIV in SSA. © 2022, The Author(s).Funding text 1: S Afzal acknowledges support of the Pakistan Society of Medical Infectious Diseases and King Edward Medical University to access the relevant data of HIV from various sources. T W Bärnighausen was supported by the Alexander von Humboldt Foundation through the Alexander von Humboldt Professor award, funded by the German Federal Ministry of Education and Research. F Carvalho and E Fernandes acknowledge support from Fundação para a Ciência e a Tecnologia (FCT), I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences - UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy - i4HB; FCT/MCTES (Ministério da Ciência, Tecnologia e Ensino Superior) through the project UIDB/50006/2020. K Deribe acknowledges support by the Wellcome Trust [grant number 201900/Z/16/Z] as part of his International Intermediate Fellowship. C Herteliu and A Pana are partially supported by a grant of the Romanian National Authority for Scientific Research and Innovation, CNDS-UEFISCDI, project number PN-III-P4-ID-PCCF-2016-0084. Claudiu Herteliu is partially supported by a grant of the Romanian Ministry of Research Innovation and Digitalization, MCID, project number ID-585-CTR-42-PFE-2021. Y J Kim acknowledges support by the Research Management Centre, Xiamen University Malaysia [No. XMUMRF/2020-C6/ITCM/0004]. S L Koulmane Laxminarayana acknowledges institutional support by the Manipal Academy of Higher Education. K Krishan acknowledges non-financial support from UGC Centre of Advanced Study, CAS II, Department of Anthropology, Panjab University, Chandigarh, India. M Kumar would like to acknowledge NIH/FIC K43 TW010716-04. I Landires is a member of the Sistema Nacional de Investigación (SNI), supported by the Secretaría Nacional de Ciencia, Tecnología e Innovación (SENACYT), Panama. V Nuñez-Samudio is a member of the Sistema Nacional de Investigación (SNI), which is supported by Panama’s Secretaría Nacional de Ciencia, Tecnología e Innovación (SENACYT). O O Odukoya was supported by the Fogarty International Center of the National Institutes of Health under the Award Number K43TW010704. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Z Quazi Syed acknowledges support from JNMC, Datta Meghe Institute of Medical Sciences. A I Ribeiro was supported by National Funds through FCT, under the ‘Stimulus of Scientific Employment – Individual Support’ program within the contract CEECIND/02386/2018. A M Samy acknowledges the support from a fellowship of the Egyptian Fulbright Mission program and Ain Shams University. R Shrestha acknowledges support from NIDA K01 Award: K01DA051346. N Taveira acknowledges support from FCT and Aga Khan Development Network (AKDN) - Portugal Collaborative Research Network in Portuguese speaking countries in Africa (project reference: 332821690), and by the European & Developing Countries Clinical Trials Partnership (EDCTP), UE (project reference: RIA2016MC-1615). B Unnikrishnan acknowledges support from Kasturba Medical College, Mangalore, Manipal Academy of Higher Education, Manipal. ; Funding text 2: LBD sub-Saharan Africa HIV Prevalence Collaborators S Afzal acknowledges support of the Pakistan Society of Medical Infectious Diseases and King Edward Medical University to access the relevant data of HIV from various sources. T W Bärnighausen was supported by the Alexander von Humboldt Foundation through the Alexander von Humboldt Professor award, funded by the German Federal Ministry of Education and Research. F Carvalho and E Fernandes acknowledge support from Fundação para a Ciência e a Tecnologia (FCT), I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences - UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy - i4HB; FCT/MCTES (Ministério da Ciência, Tecnologia e Ensino Superior) through the project UIDB/50006/2020. K Deribe acknowledges support by the Wellcome Trust [grant number 201900/Z/16/Z] as part of his International Intermediate Fellowship. C Herteliu and A Pana are partially supported by a grant of the Romanian National Authority for Scientific Research and Innovation, CNDS-UEFISCDI, project number PN-III-P4-ID-PCCF-2016-0084. Claudiu Herteliu is partially supported by a grant of the Romanian Ministry of Research Innovation and Digitalization, MCID, project number ID-585-CTR-42-PFE-2021. Y J Kim acknowledges support by the Research Management Centre, Xiamen University Malaysia [No. XMUMRF/2020-C6/ITCM/0004]. S L Koulmane Laxminarayana acknowledges institutional support by the Manipal Academy of Higher Education. K Krishan acknowledges non-financial support from UGC Centre of Advanced Study, CAS II, Department of Anthropology, Panjab University, Chandigarh, India. M Kumar would like to acknowledge NIH/FIC K43 TW010716-04. I Landires is a member of the Sistema Nacional de Investigación (SNI), supported by the Secretaría Nacional de Ciencia, Tecnología e Innovación (SENACYT), Panama. V Nuñez-Samudio is a member of the Sistema Nacional de Investigación (SNI), which is supported by Panama’s Secretaría Nacional de Ciencia, Tecnología e Innovación (SENACYT). O O Odukoya was supported by the Fogarty International Center of the National Institutes of Health under the Award Number K43TW010704. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Z Quazi Syed acknowledges support from JNMC, Datta Meghe Institute of Medical Sciences. A I Ribeiro was supported by National Funds through FCT, under the ‘Stimulus of Scientific Employment – Individual Support’ program within the contract CEECIND/02386/2018. A M Samy acknowledges the support from a fellowship of the Egyptian Fulbright Mission program and Ain Shams University. R Shrestha acknowledges support from NIDA K01 Award: K01DA051346. N Taveira acknowledges support from FCT and Aga Khan Development Network (AKDN) - Portugal Collaborative Research Network in Portuguese speaking countries in Africa (project reference: 332821690), and by the European & Developing Countries Clinical Trials Partnership (EDCTP), UE (project reference: RIA2016MC-1615). B Unnikrishnan acknowledges support from Kasturba Medical College, Mangalore, Manipal Academy of Higher Education, Manipal.; Funding text 3: This work was primarily supported by grant OPP1132415 from the Bill & Melinda Gates Foundation. The funder of the study had no role in study design, data collection, data analysis, data interpretation, writing of the report, or decision to publish. The corresponding authors had full access to all the data in the study and had final responsibility for the decision to submit for publication. ; Funding text 4: S Afzal reports leadership or fiduciary role in other board, society, committee or advocacy group, unpaid, with the Pakistan society of Community Medicine & Public Health, the Pakistan Association of Medical Editors, and the Pakistan Society of Medical Infectious Diseases, all outside the submitted work. R Ancuceanu reports 5 payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing, or educational events from Avvie, Sandoz, and B Braun, all outside the submitted work. T W Bärnighausen reports research grants from the European Union (Horizon 2020 and EIT Health), German Research Foundation (DFG), US National Institutes of Health, German Ministry of Education and Research, Alexander von Humboldt Foundation, Else-Kröner-Fresenius-Foundation, Wellcome Trust, Bill & Melinda Gates Foundation, KfW, UNAIDS, and WHO; consulting fees from KfW on the OSCAR initiative in Vietnam; participation on a Data Safety Monitoring Board or Advisory Board with the NIH-funded study “Healthy Options” (PIs: Smith Fawzi, Kaaya), Chair, Data Safety and Monitoring Board (DSMB), German National Committee on the “Future of Public Health Research and Education,” Chair of the scientific advisory board to the EDCTP Evaluation, Member of the UNAIDS Evaluation Expert Advisory Committee, National Institutes of Health Study Section Member on Population and Public Health Approaches to HIV/AIDS (PPAH), US National Academies of Sciences, Engineering, and Medicine’s Committee for the “Evaluation of Human Resources for Health in the Republic of Rwanda under the President’s Emergency Plan for AIDS Relief (PEPFAR),” University of Pennsylvania (UPenn) Population Aging Research Center (PARC) External Advisory Board Member; leadership or fiduciary role in other board, society, committee or advocacy group, paid or unpaid, as co-chair of the Global Health Hub Germany (which was initiated by the German Ministry of Health); all outside the submitted work. J das Neves reports grants or contracts from Ref. 13605 – Programa GÉNESE, Gilead Portugal (PGG/002/2016 – Programa GÉNESE, Gilead Portugal) outside the submitted work. L Dwyer-Lindgren reports support for the present manuscript from the Bill & Melinda Gates Foundation through grant OPP1132415. I Filip reports other financial or non-financial interests from Avicenna Medical and Clinical Research Institute, outside the submitted work. E Haeuser reports support for the present manuscript from the Bill & Melinda Gates Foundation through grant OPP1132415. C Herteliu reports grants from Romanian Ministry of Research Innovation and Digitalization, MCID, for project number ID-585-CTR-42-PFE-2021 (Jan 2022-Jun 2023) “Enhancing institutional performance through development of infrastructure and transdisciplinary research ecosystem within socio-economic domain – PERFECTIS,” from Romanian National Authority for Scientific Research and Innovation, CNDS-UEFISCDI, for project number PN-III-P4-ID-PCCF-2016-0084 (Oct 2018-Sep 2022) “Understanding and modelling time-space patterns of psychology-related inequalities and polarization,” and project number PN-III-P2-2.1-SOL-2020-2-0351 (Jun 2020-Oct 2020) “Approaches within public health management in the context of COVID-19 pandemic,” and from the Ministry of Labour and Social Justice, Romania for project number “Agenda for skills Romania 2020-2025”; all outside the submitted work. J J Jozwiak reports payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing, or educational events from Teva, Amgen, Synexus, Boehringer Ingelheim, Zentiva, and Sanofi as personal fees, all outside the submitted work. J Khubchandani reports other financial interests from Teva Pharmaceuticals, all outside the submitted work. K Krishnan reports other non-financial support from UGC Centre of Advanced Study, CAS II, Department of Anthropology, Panjab University, Chandigarh, India, outside the submitted work. H J Larson reports grants or contracts from the MacArthur Foundation and Merck to London School of Hygeine and Tropical Medicine, and from the Vaccine Confidence Fund to the University of Washington; payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing, or educational events from Center for Strategic and International Studies as payment to LSHTM for co-chairing HighLevel Panel and from GSK as personal payment for developing training sessions and lectures; leadership or fiduciary role in other board, society, committee or advocacy group, pair, with the ApiJect Advisory Board; all outside the submitted work. O O Odukoya reports support for the present manuscript from the Fogarty International Center of the National Institutes of Health under the Award Number K43TW010704. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. A Pans reports grants from Romanian National Authority for Scientific Research and Innovation, CNDS-UEFISCDI, for project number PN-III-P4-ID-PCCF-2016-0084 (Oct 2018-Sep 2022) “Understanding and modelling time-space patterns of psychology-related inequalities and polarization,” and project number PN-III-P2-2.1-SOL-2020-2-0351 (Jun 2020-Oct 2020) “Approaches within public health management in the context of COVID-19 pandemic,” outside the submitted work. S R Pandi-Perumal reports royalties from Springer for editing services; stock or stock options in Somnogen Canada Inc as the President and Chief Executive Officer; all outside the submitted work. A Radfar reports other financial or non-financial interests from Avicenna Medical and Clinical Research Institute, outside the submitted work. A I Ribeiro reports grants or contracts from National Funds through FCT, under the ‘Stimulus of Scientific Employment – Individual Support’ program within the contract CEECIND/02386/2018, outside the submitted work. J M Ross reports support for the present manuscript from the Bill & Melinda Gates Foundation through grant OPP1132415; grants or contracts from National Institutes of Health and Firland Foundation as payments to their institution; consulting fees from United States Agency for International Development as personal payments, and from KNCV Tuberculosis Foundation as payments to their institution; all outside the submitted work. E Rubagotti reports payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing, or educational events from the Greenwich China Office and Unviersity Prince Mohammad VI, Morocco, all outside the submitted work. B Sartorius reports grants or contracts from DHSC – GRAM Project; Leadership or fiduciary role in other board, society, committee or advocacy group, paid or unpaid, as a member of the GBD Scientific Council and a Member of WHO RGHS; all outside the submitted work. J A Singh reports consulting fees from Crealta/Horizon, Medisys, Fidia, PK Med, Two labs Inc, Adept Field Solutions, Clinical Care options, Clearview healthcare partners, Putnam associates, Focus forward, Navigant consulting, Spherix, MedIQ, Jupiter Life Science LLC, UBM LLC, Trio Health, Medscape, WebMD, and Practice Point communications, and the National Institutes of Health and the American College of Rheumatology; payment or honoraria for participating in the speakers bureau for Simply Speaking; support for attending meetings and/or travel from the steering committee of OMERACT, to attend their meeting every 2 years; participation on a Data Safety Monitoring Board or Advisory Board as an unpaid member of the FDA Arthritis Advisory Committee; leadership or fiduciary role in other board, society, committee or advocacy group, paid or unpaid, as a member of the steering committee of OMERACT, an international organization that develops measures for clinical trials and receives arm’s length funding from 12 pharmaceutical companies, with the Veterans Affairs Rheumatology Field Advisory Committee as Chair, and with the UAB Cochrane Musculoskeletal Group Satellite Center on Network Meta-analysis as a director and editor; stock or stock options in TPT Global Tech, Vaxart pharmaceuticals, Atyu Biopharma, Adaptimmune Therapeutics, GeoVax Labs, Pieris Pharmaceuticals, Enzolytics Inc, Series Therapeutics, Tonix Pharmaceuticals, and Charlotte’s Web Holdings Inc. and previously owned stock options in Amarin, Viking, and Moderna pharmaceuticals; all outside the submitted work. N Taveira reports grants or contracts from FCT and Aga Khan Development Network (AKDN) – Portugal Collaborative Research Network in Portuguese speaking countries in Africa (Project reference: 332821690) and from European & Developing Countries Clinical Trials Partnership (EDCTP), UE (Project reference: RIA2016MC-1615), as payments made to their institution, all outside the submitted work

Approaches in biotechnological applications of natural polymers

Natural polymers, such as gums and mucilage, are biocompatible, cheap, easily available and non-toxic materials of native origin. These polymers are increasingly preferred over synthetic materials for industrial applications due to their intrinsic properties, as well as they are considered alternative sources of raw materials since they present characteristics of sustainability, biodegradability and biosafety. As definition, gums and mucilages are polysaccharides or complex carbohydrates consisting of one or more monosaccharides or their derivatives linked in bewildering variety of linkages and structures. Natural gums are considered polysaccharides naturally occurring in varieties of plant seeds and exudates, tree or shrub exudates, seaweed extracts, fungi, bacteria, and animal sources. Water-soluble gums, also known as hydrocolloids, are considered exudates and are pathological products; therefore, they do not form a part of cell wall. On the other hand, mucilages are part of cell and physiological products. It is important to highlight that gums represent the largest amounts of polymer materials derived from plants. Gums have enormously large and broad applications in both food and non-food industries, being commonly used as thickening, binding, emulsifying, suspending, stabilizing agents and matrices for drug release in pharmaceutical and cosmetic industries. In the food industry, their gelling properties and the ability to mold edible films and coatings are extensively studied. The use of gums depends on the intrinsic properties that they provide, often at costs below those of synthetic polymers. For upgrading the value of gums, they are being processed into various forms, including the most recent nanomaterials, for various biotechnological applications. Thus, the main natural polymers including galactomannans, cellulose, chitin, agar, carrageenan, alginate, cashew gum, pectin and starch, in addition to the current researches about them are reviewed in this article.. }To the Conselho Nacional de Desenvolvimento Cientfíico e Tecnológico (CNPq) for fellowships (LCBBC and MGCC) and the Coordenação de Aperfeiçoamento de Pessoal de Nvíel Superior (CAPES) (PBSA). This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit, the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462) and COMPETE 2020 (POCI-01-0145-FEDER-006684) (JAT)

DMTs and Covid-19 severity in MS: a pooled analysis from Italy and France

We evaluated the effect of DMTs on Covid-19 severity in patients with MS, with a pooled-analysis of two large cohorts from Italy and France. The association of baseline characteristics and DMTs with Covid-19 severity was assessed by multivariate ordinal-logistic models and pooled by a fixed-effect meta-analysis. 1066 patients with MS from Italy and 721 from France were included. In the multivariate model, anti-CD20 therapies were significantly associated (OR = 2.05, 95%CI = 1.39–3.02, p < 0.001) with Covid-19 severity, whereas interferon indicated a decreased risk (OR = 0.42, 95%CI = 0.18–0.99, p = 0.047). This pooled-analysis confirms an increased risk of severe Covid-19 in patients on anti-CD20 therapies and supports the protective role of interferon

Mapping geographical inequalities in oral rehydration therapy coverage in low-income and middle-income countries, 2000-17

Background Oral rehydration solution (ORS) is a form of oral rehydration therapy (ORT) for diarrhoea that has the potential to drastically reduce child mortality; yet, according to UNICEF estimates, less than half of children younger than 5 years with diarrhoea in low-income and middle-income countries (LMICs) received ORS in 2016. A variety of recommended home fluids (RHF) exist as alternative forms of ORT; however, it is unclear whether RHF prevent child mortality. Previous studies have shown considerable variation between countries in ORS and RHF use, but subnational variation is unknown. This study aims to produce high-resolution geospatial estimates of relative and absolute coverage of ORS, RHF, and ORT (use of either ORS or RHF) in LMICs. Methods We used a Bayesian geostatistical model including 15 spatial covariates and data from 385 household surveys across 94 LMICs to estimate annual proportions of children younger than 5 years of age with diarrhoea who received ORS or RHF (or both) on continuous continent-wide surfaces in 2000-17, and aggregated results to policy-relevant administrative units. Additionally, we analysed geographical inequality in coverage across administrative units and estimated the number of diarrhoeal deaths averted by increased coverage over the study period. Uncertainty in the mean coverage estimates was calculated by taking 250 draws from the posterior joint distribution of the model and creating uncertainty intervals (UIs) with the 2 center dot 5th and 97 center dot 5th percentiles of those 250 draws. Findings While ORS use among children with diarrhoea increased in some countries from 2000 to 2017, coverage remained below 50% in the majority (62 center dot 6%; 12 417 of 19 823) of second administrative-level units and an estimated 6 519 000 children (95% UI 5 254 000-7 733 000) with diarrhoea were not treated with any form of ORT in 2017. Increases in ORS use corresponded with declines in RHF in many locations, resulting in relatively constant overall ORT coverage from 2000 to 2017. Although ORS was uniformly distributed subnationally in some countries, within-country geographical inequalities persisted in others; 11 countries had at least a 50% difference in one of their units compared with the country mean. Increases in ORS use over time were correlated with declines in RHF use and in diarrhoeal mortality in many locations, and an estimated 52 230 diarrhoeal deaths (36 910-68 860) were averted by scaling up of ORS coverage between 2000 and 2017. Finally, we identified key subnational areas in Colombia, Nigeria, and Sudan as examples of where diarrhoeal mortality remains higher than average, while ORS coverage remains lower than average. Interpretation To our knowledge, this study is the first to produce and map subnational estimates of ORS, RHF, and ORT coverage and attributable child diarrhoeal deaths across LMICs from 2000 to 2017, allowing for tracking progress over time. Our novel results, combined with detailed subnational estimates of diarrhoeal morbidity and mortality, can support subnational needs assessments aimed at furthering policy makers' understanding of within-country disparities. Over 50 years after the discovery that led to this simple, cheap, and life-saving therapy, large gains in reducing mortality could still be made by reducing geographical inequalities in ORS coverage. Copyright (c) 2020 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license.Peer reviewe

Five insights from the Global Burden of Disease Study 2019

The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019 provides a rules-based synthesis of the available evidence on levels and trends in health outcomes, a diverse set of risk factors, and health system responses. GBD 2019 covered 204 countries and territories, as well as first administrative level disaggregations for 22 countries, from 1990 to 2019. Because GBD is highly standardised and comprehensive, spanning both fatal and non-fatal outcomes, and uses a mutually exclusive and collectively exhaustive list of hierarchical disease and injury causes, the study provides a powerful basis for detailed and broad insights on global health trends and emerging challenges. GBD 2019 incorporates data from 281 586 sources and provides more than 3.5 billion estimates of health outcome and health system measures of interest for global, national, and subnational policy dialogue. All GBD estimates are publicly available and adhere to the Guidelines on Accurate and Transparent Health Estimate Reporting. From this vast amount of information, five key insights that are important for health, social, and economic development strategies have been distilled. These insights are subject to the many limitations outlined in each of the component GBD capstone papers.Peer reviewe