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      Inhibition of Severe Acute Respiratory Syndrome Coronavirus 2 Replication by Hypertonic Saline Solution in Lung and Kidney Epithelial Cells

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          Abstract

          An unprecedented global health crisis has been caused by a new virus called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We performed experiments to test if a hypertonic saline solution was capable of inhibiting virus replication. Our data show that 1.2% NaCl inhibited virus replication by 90%, achieving 100% of inhibition at 1.5% in the nonhuman primate kidney cell line Vero, and 1.1% of NaCl was sufficient to inhibit the virus replication by 88% in human epithelial lung cell line Calu-3. Furthermore, our results indicate that the inhibition is due to an intracellular mechanism and not to the dissociation of the spike SARS-CoV-2 protein and its human receptor. NaCl depolarizes the plasma membrane causing a low energy state (high ADP/ATP concentration ratio) without impairing mitochondrial function, supposedly associated with the inhibition of the SARS-CoV-2 life cycle. Membrane depolarization and intracellular energy deprivation are possible mechanisms by which the hypertonic saline solution efficiently prevents virus replication in vitro assays.

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          Most cited references67

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          A Novel Coronavirus from Patients with Pneumonia in China, 2019

          Summary In December 2019, a cluster of patients with pneumonia of unknown cause was linked to a seafood wholesale market in Wuhan, China. A previously unknown betacoronavirus was discovered through the use of unbiased sequencing in samples from patients with pneumonia. Human airway epithelial cells were used to isolate a novel coronavirus, named 2019-nCoV, which formed a clade within the subgenus sarbecovirus, Orthocoronavirinae subfamily. Different from both MERS-CoV and SARS-CoV, 2019-nCoV is the seventh member of the family of coronaviruses that infect humans. Enhanced surveillance and further investigation are ongoing. (Funded by the National Key Research and Development Program of China and the National Major Project for Control and Prevention of Infectious Disease in China.)
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            SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor

            Summary The recent emergence of the novel, pathogenic SARS-coronavirus 2 (SARS-CoV-2) in China and its rapid national and international spread pose a global health emergency. Cell entry of coronaviruses depends on binding of the viral spike (S) proteins to cellular receptors and on S protein priming by host cell proteases. Unravelling which cellular factors are used by SARS-CoV-2 for entry might provide insights into viral transmission and reveal therapeutic targets. Here, we demonstrate that SARS-CoV-2 uses the SARS-CoV receptor ACE2 for entry and the serine protease TMPRSS2 for S protein priming. A TMPRSS2 inhibitor approved for clinical use blocked entry and might constitute a treatment option. Finally, we show that the sera from convalescent SARS patients cross-neutralized SARS-2-S-driven entry. Our results reveal important commonalities between SARS-CoV-2 and SARS-CoV infection and identify a potential target for antiviral intervention.
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              Is Open Access

              Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation

              Structure of the nCoV trimeric spike The World Health Organization has declared the outbreak of a novel coronavirus (2019-nCoV) to be a public health emergency of international concern. The virus binds to host cells through its trimeric spike glycoprotein, making this protein a key target for potential therapies and diagnostics. Wrapp et al. determined a 3.5-angstrom-resolution structure of the 2019-nCoV trimeric spike protein by cryo–electron microscopy. Using biophysical assays, the authors show that this protein binds at least 10 times more tightly than the corresponding spike protein of severe acute respiratory syndrome (SARS)–CoV to their common host cell receptor. They also tested three antibodies known to bind to the SARS-CoV spike protein but did not detect binding to the 2019-nCoV spike protein. These studies provide valuable information to guide the development of medical counter-measures for 2019-nCoV. Science, this issue p. 1260
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                Author and article information

                Journal
                ACS Pharmacol Transl Sci
                ACS Pharmacol Transl Sci
                pt
                aptsfn
                ACS Pharmacology & Translational Science
                American Chemical Society
                2575-9108
                03 September 2021
                : acsptsci.1c00080
                Affiliations
                []Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo , São Paulo 05508, Brazil
                []Department of Biochemistry, Institute of Chemistry, University of São Paulo , São Paulo 05508, Brazil
                [§ ]Medical School Clinical Hospital, University of São Paulo , São Paulo 05508, Brazil
                []Gaspar Vianna Clinic Hospital Foundation , Belém 66083, Brazil
                []Hospital Israelita Albert Einstein , São Paulo 05652, Brazil
                [# ]LIM-03, Central Laboratories Division, Clinics Hospital, São Paulo School of Medicine, University of São Paulo , São Paulo 05508, Brazil
                []LIM-07, Institute of Tropical Medicine, Department of Gastroenterology, University of São Paulo School of Medicine , São Paulo 05508, Brazil
                []Scientific Platform Pasteur USP , São Paulo 05508, Brazil
                []Development and Innovation Center, Laboratory of Virology, Butantan Institute , São Paulo 05503, Brazil
                Author notes
                Author information
                https://orcid.org/0000-0002-2114-3815
                https://orcid.org/0000-0002-5664-8055
                Article
                10.1021/acsptsci.1c00080
                8442612
                34651104
                dc6ad333-655f-43ef-98ef-b6e915cd8a82
                © 2021 American Chemical Society

                This article is made available via the PMC Open Access Subset for unrestricted RESEARCH re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

                History
                : 15 March 2021
                Funding
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo, doi 10.13039/501100001807;
                Award ID: 2015/13345-1
                Funded by: Financiadora de Estudos e Projetos, doi 10.13039/501100004809;
                Award ID: FINEP 0459/20
                Funded by: Conselho Nacional de Desenvolvimento Científico e Tecnológico, doi 10.13039/501100003593;
                Award ID: 465656/2014-5
                Funded by: Conselho Nacional de Desenvolvimento Científico e Tecnológico, doi 10.13039/501100003593;
                Award ID: 306392/2017-8
                Funded by: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, doi 10.13039/501100002322;
                Award ID: 88887.131387/2016-00
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo, doi 10.13039/501100001807;
                Award ID: 2020/06409-1
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo, doi 10.13039/501100001807;
                Award ID: 2020/04680-0
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo, doi 10.13039/501100001807;
                Award ID: 2019/00195-2
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo, doi 10.13039/501100001807;
                Award ID: 2018/07366-4
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo, doi 10.13039/501100001807;
                Award ID: 2017/24769-2
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo, doi 10.13039/501100001807;
                Award ID: 2016/20045-7
                Categories
                Article
                Custom metadata
                pt1c00080
                pt1c00080

                sars-cov-2,nacl,virus inhibition,cell depolarization,atp
                sars-cov-2, nacl, virus inhibition, cell depolarization, atp

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