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1.
Trends Biotechnol ; 40(8): 987-1003, 2022 Aug.
Article in English | MEDLINE | ID: covidwho-1931129

ABSTRACT

Industry 4.0 encompasses a new industrial revolution in which advanced manufacturing systems are interconnected with information technologies. These sophisticated data-gathering technologies have led to a shift toward smarter manufacturing processes involving the use of smart materials (SMs). The properties of SMs make them highly attractive for numerous biomedical applications. The integration of artificial intelligence (AI) enables them to be effectively used in the design of novel biomedical platforms to overcome shortcomings in the current biotechnology industry. This review summarizes recent advances in AI-assisted SMs for different healthcare products. The current challenges and future perspectives of AI-supported smart biosystems are also discussed, particularly with the regard to their applications in drug design, biosensors, theranostics, and electronic skins.


Subject(s)
Biosensing Techniques , Wearable Electronic Devices , Artificial Intelligence , Biotechnology , Precision Medicine
2.
Proc Natl Acad Sci U S A ; 119(28): e2118260119, 2022 Jul 12.
Article in English | MEDLINE | ID: covidwho-1908380

ABSTRACT

Type VI CRISPR-Cas systems have been repurposed for various applications such as gene knockdown, viral interference, and diagnostics. However, the identification and characterization of thermophilic orthologs will expand and unlock the potential of diverse biotechnological applications. Herein, we identified and characterized a thermostable ortholog of the Cas13a family from the thermophilic organism Thermoclostridium caenicola (TccCas13a). We show that TccCas13a has a close phylogenetic relation to the HheCas13a ortholog from the thermophilic bacterium Herbinix hemicellulosilytica and shares several properties such as thermostability and inability to process its own pre-CRISPR RNA. We demonstrate that TccCas13a possesses robust cis and trans activities at a broad temperature range of 37 to 70 °C, compared with HheCas13a, which has a more limited range and lower activity. We harnessed TccCas13a thermostability to develop a sensitive, robust, rapid, and one-pot assay, named OPTIMA-dx, for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection. OPTIMA-dx exhibits no cross-reactivity with other viruses and a limit of detection of 10 copies/µL when using a synthetic SARS-CoV-2 genome. We used OPTIMA-dx for SARS-CoV-2 detection in clinical samples, and our assay showed 95% sensitivity and 100% specificity compared with qRT-PCR. Furthermore, we demonstrated that OPTIMA-dx is suitable for multiplexed detection and is compatible with the quick extraction protocol. OPTIMA-dx exhibits critical features that enable its use at point of care (POC). Therefore, we developed a mobile phone application to facilitate OPTIMA-dx data collection and sharing of patient sample results. This work demonstrates the power of CRISPR-Cas13 thermostable enzymes in enabling key applications in one-pot POC diagnostics and potentially in transcriptome engineering, editing, and therapies.


Subject(s)
Bacterial Proteins , COVID-19 , CRISPR-Associated Proteins , Clostridiales , Endodeoxyribonucleases , Point-of-Care Testing , SARS-CoV-2 , Bacterial Proteins/chemistry , Bacterial Proteins/classification , Bacterial Proteins/genetics , Biotechnology , COVID-19/diagnosis , CRISPR-Associated Proteins/chemistry , CRISPR-Associated Proteins/classification , CRISPR-Associated Proteins/genetics , Clostridiales/enzymology , Endodeoxyribonucleases/chemistry , Endodeoxyribonucleases/classification , Endodeoxyribonucleases/genetics , Enzyme Stability , Hot Temperature , Humans , Phylogeny , SARS-CoV-2/isolation & purification
4.
Cad. Saúde Pública (Online) ; 37(4): e00254720, 2021. graf
Article in Portuguese | WHO COVID, LILACS (Americas) | ID: covidwho-1862330

ABSTRACT

Resumo: Em tempos de pandemia, a vulnerabilidade econômica, tecnológica e dos sistemas de saúde fica ainda mais exposta. No Brasil, os maiores desafios são o controle do déficit da balança comercial e a dificuldade de acesso a medicamentos e produtos da saúde ou até mesmo de seu desenvolvimento. A forte dependência externa de insumos e produtos para a saúde é um dos fatores negativos do país no enfrentamento da emergência sanitária mundial ocasionada pelo novo coronavírus SARS-CoV-2. Nesse contexto, o artigo procurou discutir as vulnerabilidades do subsistema de base química e biotecnológica nacional diante da atual pandemia, tendo como foco a indústria de medicamentos e de produtos biotecnológicos e a articulação entre os campos da saúde e da economia política. Realizou-se pesquisa qualitativa, utilizando, como procedimentos metodológicos, revisão bibliográfica e análise de dados secundários. Os resultados encontrados, relacionados a baixo investimento em saúde pública, baixa capacidade de inovação, perfil da indústria farmacêutica, dependência externa, política cambial, patentes, entre outros, demonstram a grande fragilidade inovativa e tecnológica da indústria de medicamentos e produtos biotecnológicos e a essencialidade de articulação entre diversos campos, em especial, da saúde e da economia, para incorporação de uma visão sistêmica, que crie condições para redução das vulnerabilidades, no que tange ao enfrentamento da pandemia, e equacione um projeto de desenvolvimento para o país.


Abstract: During a pandemic, economic, technological, and health systems' vulnerability become even more evident. A key challenge in Brazil is to control the trade deficit and difficulty in access to medicines and health products, even their development. Brazil's heavy external dependency on health inputs and products is one of the negative factors in confronting the global health emergency caused by SARS-CoV-2. The article aimed to discuss the vulnerabilities of the domestic chemical and biotechnological subsystem in the face of the current pandemic, with a focus on the pharmaceutical and biotech industry and the linkage between the fields of health and political economics. A qualitative study was performed with a literature review and analysis of secondary data as the methodological procedures. The results revealed low investment in public health, low innovation capacity, the pharmaceutical industry's profile, external dependency, currency exchange policy, patents, and other factors, demonstrating the major vulnerability in innovation and technology in the domestic pharmaceutical and biotech industry and the essential nature of linkage between various fields, especially health and the economy, for the incorporation of a systemic vision that creates the conditions to reduce vulnerabilities in the response to the pandemic and promote a development project for the country.


Resumen: En tiempos de pandemia, la vulnerabilidad económica, tecnológica y de los sistemas de salud queda aún más expuesta. En Brasil, uno de los mayores desafíos es el control del déficit de la balanza comercial, así como la dificultad de acceso a medicamentos y produtos de salud o incluso de su desarrollo. La fuerte dependencia externa de insumos y productos para la salud es uno de los factores negativos del país en el combate a la emergencia sanitaria mundial, ocasionada por el nuevo coronavirus SARS-CoV-2. En este contexto, el artículo procuró discutir las vulnerabilidades del subsistema de base química y biotecnológica nacional ante la actual pandemia, poniendo el foco en la industria de medicamentos y de productos biotecnológicos, así como la coordinación entre los campos de la salud y economía política. Se realizó una investigación cualitativa, utilizando como procedimientos metodológicos la revisión bibliográfica y el análisis de datos secundarios. Los resultados hallados, relacionados con la baja inversión en salud pública, baja capacidad de innovación, perfil de la industria farmacéutica, dependencia externa, política de cambio, patentes, entre otros, demuestra la gran fragilidad innovadora y tecnológica de la industria de medicamentos, así como de productos biotecnológicos, y la necesidad de una coordinación entre diversas áreas, en especial, de la salud y economía, para que se incorpore una visión sistémica, que cree condiciones para la reducción de las vulnerabilidades, en lo que atañe al combate de la pandemia y que cree un proyecto equilibrado de desarrollo para el país.


Subject(s)
Humans , Pharmaceutical Preparations , COVID-19 , Biotechnology , Brazil/epidemiology , Pandemics , SARS-CoV-2
5.
Lab Chip ; 22(8): 1469-1473, 2022 04 12.
Article in English | MEDLINE | ID: covidwho-1805673

ABSTRACT

The COVID-19 pandemic has proven the need for point-of-care diagnosis of respiratory diseases and microfluidic technology has risen to the occasion. Mesa Biotech (San Diego, CA) originally developed the Accula platform for the diagnosis of influenza A and B and then extended the platform to SARS-CoV-2. Mesa Biotech has experienced tremendous success, culminating in acquisition by Thermo Fisher for up to $550m USD. The Accula microfluidics platform accomplished the leap from the lab to commercial product through clever design and engineering choices. Through information obtained from interviews with key Mesa Biotech leaders and publicly-available documents, we describe the keys to Mesa's success and how they might inform other lab-on-a-chip companies.


Subject(s)
COVID-19 , Pandemics , Biotechnology , COVID-19/diagnosis , Humans , Microfluidics , SARS-CoV-2
6.
Cells ; 11(7)2022 03 31.
Article in English | MEDLINE | ID: covidwho-1776138

ABSTRACT

The outbreak of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause for the ongoing global public health emergency. It is more commonly known as coronavirus disease 2019 (COVID-19); the pandemic threat continues to spread aroundthe world with the fluctuating emergence of its new variants. The severity of COVID-19 ranges from asymptomatic to serious acute respiratory distress syndrome (ARDS), which has led to a high human mortality rate and disruption of socioeconomic well-being. For the restoration of pre-pandemic normalcy, the international scientific community has been conducting research on a war footing to limit extremely pathogenic COVID-19 through diagnosis, treatment, and immunization. Since the first report of COVID-19 viral infection, an array of laboratory-based and point-of-care (POC) approaches have emerged for diagnosing and understanding its status of outbreak. The RT-PCR-based viral nucleic acid test (NAT) is one of the rapidly developed and most used COVID-19 detection approaches. Notably, the current forbidding status of COVID-19 requires the development of safe, targeted vaccines/vaccine injections (shots) that can reduce its associated morbidity and mortality. Massive and accelerated vaccination campaigns would be the most effective and ultimate hope to end the COVID-19 pandemic. Since the SARS-CoV-2 virus outbreak, emerging biotechnologies and their multidisciplinary approaches have accelerated the understanding of molecular details as well as the development of a wide range of diagnostics and potential vaccine candidates, which are indispensable to combating the highly contagious COVID-19. Several vaccine candidates have completed phase III clinical studies and are reported to be effective in immunizing against COVID-19 after their rollout via emergency use authorization (EUA). However, optimizing the type of vaccine candidates and its route of delivery that works best to control viral spread is crucial to face the threatening variants expected to emerge over time. In conclusion, the insights of this review would facilitate the development of more likely diagnostics and ideal vaccines for the global control of COVID-19.


Subject(s)
COVID-19 , Pandemics , Biotechnology , COVID-19/diagnosis , COVID-19/epidemiology , COVID-19/prevention & control , COVID-19 Vaccines , Humans , Pandemics/prevention & control , SARS-CoV-2
7.
Technol Health Care ; 30(2): 513-516, 2022.
Article in English | MEDLINE | ID: covidwho-1753336

ABSTRACT

BACKGROUND: Technologies for COVID-19 are in high demand, and supply chains from biotech and pharma industries face critical supply issues. OBJECTIVE: This paper discusses the case on mRNA technologies and quality assurance issues. METHODS: Interviews with managers of biotech companies were performed. These helped to prepare biotech panels at a roundtable on quality of medicine, organized by the Polish Academy of Sciences in Paris (PAN) in 2021. RESULTS: This paper analyzes the new mRNA technology and shows the importance of emerging new biotech firms, especially tool companies providing services in quality assurance. It highlights research areas and types of survey instruments with academic collaborators, to better understand the economics of mRNA technology platforms. CONCLUSIONS: A future research agenda for collaboration with academic communities is proposed on cost-quality tradeoffs and clinical quality attributes for such genomic technologies.


Subject(s)
COVID-19 , Biotechnology , COVID-19/epidemiology , Humans , Industry
8.
Proc Natl Acad Sci U S A ; 119(14): e2119093119, 2022 04 05.
Article in English | MEDLINE | ID: covidwho-1751830

ABSTRACT

SignificanceUsing SARS-CoV-2 as a relevant case study for infectious disease, we investigate the structure-function relationships that dictate antiviral spherical nucleic acid (SNA) vaccine efficacy. We show that the SNA architecture can be rapidly employed to target COVID-19 through incorporation of the receptor-binding domain, and that the resulting vaccine potently activates human cells in vitro and mice in vivo. Furthermore, when challenged with a lethal viral infection, only mice treated with the SNA vaccine survived. Taken together, this work underscores the importance of rational vaccine design for infectious disease to yield vaccines that elicit more potent immune responses to effectively fight disease.


Subject(s)
Communicable Disease Control , Nucleic Acids/immunology , Vaccines, DNA/immunology , Animals , Biotechnology , COVID-19/prevention & control , Communicable Disease Control/methods , Communicable Diseases/etiology , Communicable Diseases/immunology , Humans , Nucleic Acids/chemistry , SARS-CoV-2/immunology , Vaccine Development , Vaccines, DNA/genetics , Viral Vaccines/genetics , Viral Vaccines/immunology
9.
Int J Environ Res Public Health ; 19(4)2022 02 18.
Article in English | MEDLINE | ID: covidwho-1701031

ABSTRACT

Several coronaviruses (CoVs) have been identified as human pathogens, including the α-CoVs strains HCoV-229E and HCoV-NL63 and the ß-CoVs strains HCoV-HKU1 and HCoV-OC43. SARS-CoV, MERS-CoV, and SARS-CoV-2 are also classified as ß-coronavirus. New SARS-CoV-2 spike genomic variants are responsible for human-to-human and interspecies transmissibility, consequences of adaptations of strains from animals to humans. The receptor-binding domain (RBD) of SARS-CoV-2 binds to receptor ACE2 in humans and animal species with high affinity, suggesting there have been adaptive genomic variants. New genomic variants including the incorporation, replacement, or deletion of the amino acids at a variety of positions in the S protein have been documented and are associated with the emergence of new strains adapted to different hosts. Interactions between mutated residues and RBD have been demonstrated by structural modelling of variants including D614G, B.1.1.7, B1.351, P.1, P2; other genomic variants allow escape from antibodies generated by vaccines. Epidemiological and molecular tools are being used for real-time tracking of pathogen evolution and particularly new SARS-CoV-2 variants. COVID-19 vaccines obtained from classical and next-generation vaccine production platforms have entered clinicals trials. Biotechnology strategies of the first generation (attenuated and inactivated virus-CoronaVac, CoVaxin; BBIBP-CorV), second generation (replicating-incompetent vector vaccines-ChAdOx-1; Ad5-nCoV; Sputnik V; JNJ-78436735 vaccine-replicating-competent vector, protein subunits, virus-like particles-NVX-CoV2373 vaccine), and third generation (nucleic-acid vaccines-INO-4800 (DNA); mRNA-1273 and BNT 162b (RNA vaccines) have been used. Additionally, dendritic cells (LV-SMENP-DC) and artificial antigen-presenting (aAPC) cells modified with lentiviral vector have also been developed to inhibit viral activity. Recombinant vaccines against COVID-19 are continuously being applied, and new clinical trials have been tested by interchangeability studies of viral vaccines developed by classical and next-generation platforms.


Subject(s)
COVID-19 Vaccines , COVID-19 , Ad26COVS1 , Animals , Biotechnology , COVID-19/prevention & control , Genomics , Humans , SARS-CoV-2/genetics , Spike Glycoprotein, Coronavirus/chemistry , Spike Glycoprotein, Coronavirus/genetics
10.
Nat Commun ; 13(1): 968, 2022 02 18.
Article in English | MEDLINE | ID: covidwho-1705624

ABSTRACT

DNA/RNA-gold nanoparticle (DNA/RNA-AuNP) nanoprobes have been widely employed for nanobiotechnology applications. Here, we discover that both thiolated and non-thiolated DNA/RNA can be efficiently attached to AuNPs to achieve high-stable spherical nucleic acid (SNA) within minutes under a domestic microwave (MW)-assisted heating-dry circumstance. Further studies show that for non-thiolated DNA/RNA the conjugation is poly (T/U) tag dependent. Spectroscopy, test strip hybridization, and loading counting experiments indicate that low-affinity poly (T/U) tag mediates the formation of a standing-up conformation, which is distributed in the outer layer of SNA structure. In further application studies, CRISPR/Cas9-sgRNA (136 bp), SARS-CoV-2 RNA fragment (1278 bp), and rolling circle amplification (RCA) DNA products (over 1000 bp) can be successfully attached on AuNPs, which overcomes the routine methods in long-chain nucleic acid-AuNP conjugation, exhibiting great promise in biosensing and nucleic acids delivery applications. Current heating-dry strategy has improved traditional DNA/RNA-AuNP conjugation methods in simplicity, rapidity, cost, and universality.


Subject(s)
Biosensing Techniques/methods , Gold/chemistry , Metal Nanoparticles/chemistry , Biotechnology/methods , COVID-19/diagnosis , COVID-19/virology , COVID-19 Nucleic Acid Testing/methods , DNA/chemistry , Heating/methods , Humans , Limit of Detection , Microwaves , Nanomedicine/methods , RNA, Viral/chemistry , RNA, Viral/genetics , RNA, Viral/isolation & purification , SARS-CoV-2/genetics
13.
EMBO Rep ; 22(9): e53229, 2021 09 06.
Article in English | MEDLINE | ID: covidwho-1576871

ABSTRACT

New biotechnologies such as gene drives and engineered viruses herald a viral era that would give humans exceptional power over any organism at the level of the genotype.


Subject(s)
Biotechnology , Gene Drive Technology , Humans
14.
Microb Biotechnol ; 15(1): 3-4, 2022 01.
Article in English | MEDLINE | ID: covidwho-1598718

ABSTRACT

This year, 2022, Microbial Biotechnology (MBT) celebrates its 15th birthday. In journal terms, 15 is adulthood. It has been a privilege to develop the idea, launch the journal, nurture it in its infancy and stimulate and support it during its adolescence and early adulthood. The success of MBT - its growth over the last 4 years averaging > 30%, and the highest impact factor in the field, making it the leading research journal in applied microbiology/microbial biotechnology and the most attractive to publish in - gives us enormous pleasure and satisfaction, and not a little pride.


Subject(s)
Parenting , Publishing , Biotechnology
16.
Drug Discov Today ; 26(11): 2515-2526, 2021 11.
Article in English | MEDLINE | ID: covidwho-1540581

ABSTRACT

Over the past few decades, the number of health and 'omics-related data' generated and stored has grown exponentially. Patient information can be collected in real time and explored using various artificial intelligence (AI) tools in clinical trials; mobile devices can also be used to improve aspects of both the diagnosis and treatment of diseases. In addition, AI can be used in the development of new drugs or for drug repurposing, in faster diagnosis and more efficient treatment for various diseases, as well as to identify data-driven hypotheses for scientists. In this review, we discuss how AI is starting to revolutionize the life sciences sector.


Subject(s)
Artificial Intelligence , Biological Science Disciplines , Biotechnology , Clinical Trials as Topic , Data Science , Drug Design , Drug Development , Electronic Health Records , Humans , Mobile Applications , Natural Language Processing , Pharmacology , Publishing
17.
Microb Biotechnol ; 15(1): 135-148, 2022 01.
Article in English | MEDLINE | ID: covidwho-1522635

ABSTRACT

Pfizer-BioNTech and Moderna developed in record time mRNA vaccines against COVID-19 of high efficacy. The modest protection achieved with a similarly designed mRNA from CureVac underlines the importance of biotechnological details in formulation such as replacement of uridine by pseudouridine in the mRNA encoding the SARS-CoV-2 spike protein or the lipid composition of the nanoparticle coating the mRNA. Phase 3 vaccine trials and vaccine studies in special subject groups as well observational studies in whole populations confirmed the real-world vaccine efficacy against symptomatic disease, particularly against severe COVID-19 cases and to a lesser extent against mild SARS-CoV-2 infections. mRNA vaccine protection extended also to the alpha and beta variant viruses. The surge of delta variants led to an increase of infections and cases even in populations which achieved high vaccine coverage. This efficacy decline resulted to a lesser extent from a weaker neutralization of the delta variant but mostly from a waning vaccine protection over time. Data from Israel documented the efficacy of a third 'booster' injection 5 months after the second injection in older segments of the population. Adverse reactions consisted of transient injection site pain, headache, muscle pain, fatigue, fever and chills. Extensive surveillance studies documented a good safety profile revealing only a non-significant increase in transient facial nerve paralysis and a significant, but modest increase in myocarditis in vaccinated young males that was lower than the myocarditis risk induced by SARS-CoV-2 infection.


Subject(s)
COVID-19 , Aged , Biotechnology , COVID-19 Vaccines , Humans , Male , SARS-CoV-2 , Spike Glycoprotein, Coronavirus , Vaccines, Synthetic
18.
Sheng Wu Gong Cheng Xue Bao ; 37(10): 3405-3410, 2021 Oct 25.
Article in Chinese | MEDLINE | ID: covidwho-1497978

ABSTRACT

The international cooperation project "electricity-driven low energy and chemical input technology for accelerated bioremediation" (abridged as "ELECTRA") is jointly supported by National Nature Science Foundation of China (NSFC) and European Commission (EC). The ELECTRA consortium consists of 5 research institutions and universities from China and 17 European research institutions and universities, as well as high-tech companies of EC countries. ELECTRA focuses on researches of biodegradation of emerging organic compounds (EOCs) and novel environmental biotechnologies of low-energy and low-chemical inputs. The project has been successfully operated for 2 years, and has made important progresses in obtaining EOCs-degrading microbes, developing weak-electricity-accelerated bioremediation, and 3D-printing techniques for microbial consortium. The ELECTRA has promoted collaborations among the Chinese and European scientists. In the future, ELECTRA will overcome the negative impact of the COVID-19 pandemic and fulfill the scientific objectives through strengthening the international collaboration.


Subject(s)
COVID-19 , Pandemics , Biodegradation, Environmental , Biotechnology , Electricity , Humans , SARS-CoV-2
19.
Viruses ; 13(10)2021 10 14.
Article in English | MEDLINE | ID: covidwho-1481012

ABSTRACT

Viruses may cause devastating diseases in several organisms; however, they are simple systems that can be manipulated to be beneficial and useful for many purposes in different areas. In medicine, viruses have been used for a long time in vaccines and are now being used as vectors to carry materials for the treatment of diseases, such as cancer, being able to target specific cells. In agriculture, viruses are being studied to introduce desirable characteristics in plants or render resistance to biotic and abiotic stresses. Viruses have been exploited in nanotechnology for the deposition of specific metals and have been shown to be of great benefit to nanomaterial production. They can also be used for different applications in pharmacology, cosmetics, electronics, and other industries. Thus, viruses are no longer only seen as enemies. They have shown enormous potential, covering several important areas in our lives, and they are making our lives easier and better. Although viruses have already proven their potential, there is still a long road ahead. This prompt us to propose this theme in the Special Issue "The application of viruses to biotechnology". We believe that the articles gathered here highlight recent significant advances in the use of viruses in several fields, contributing to the current knowledge on virus applications.


Subject(s)
Biotechnology , Viruses , Agriculture , Animals , Gene Expression , Genetic Therapy , Genetic Vectors , Humans , Nanostructures , Nanotechnology , Plants/virology , Viral Vaccines , Viruses/genetics
20.
Nucleic Acid Ther ; 31(5): 321-323, 2021 10.
Article in English | MEDLINE | ID: covidwho-1467290

ABSTRACT

The utilization of the mRNA-based Pfizer-BioNTech and Moderna coronavirus disease 2019 (COVID-19) vaccines represents the culmination of many years of nonviral nucleic acid delivery, but more importantly, they signify a massive clinical scientific success. Scientists working in the area of nucleic acid delivery using lipid nanoparticles will undoubtedly be energized by the success of these vaccines and begin to collect much needed data in the realm of nonviral-based RNA and DNA delivery, specifically, the use of lipid nanoparticles, the immune response, safety, and efficacy. It is easily conceivable that in the future we can utilize these data to help streamline our approach for the delivery of DNA for gene therapy and regulatory RNAs for therapeutic and regenerative medicine (ie, wound repair) applications.


Subject(s)
COVID-19 Vaccines/administration & dosage , COVID-19/prevention & control , DNA/pharmacokinetics , Gene Transfer Techniques , RNA, Messenger/pharmacokinetics , Biotechnology/trends , COVID-19/immunology , COVID-19/virology , COVID-19 Vaccines/biosynthesis , COVID-19 Vaccines/chemistry , COVID-19 Vaccines/genetics , DNA/chemistry , Data Mining , Dependovirus/genetics , Dependovirus/immunology , Humans , Liposomes/chemistry , Liposomes/pharmacokinetics , Nanoparticles/administration & dosage , Nanoparticles/chemistry , RNA, Messenger/chemistry , SARS-CoV-2/drug effects , SARS-CoV-2/immunology , SARS-CoV-2/pathogenicity
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