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Article ; Online: Propagation and Quantification of SARS-CoV-2.

Jureka, Alexander S / Basler, Christopher F

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2452, Page(s) 111–129

Abstract: In late 2019, the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, China. Since its emergence, SARS-CoV-2 has been responsible for a world-wide pandemic resulting in over 80 million infections and over 1.8 ... ...

Abstract	In late 2019, the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, China. Since its emergence, SARS-CoV-2 has been responsible for a world-wide pandemic resulting in over 80 million infections and over 1.8 million deaths. The severity of the pandemic has prompted widespread research efforts to more fully understand SARS-CoV-2 and the disease it causes, COVID-19. Research into this novel virus will be facilitated by the availability of clearly described and effective protocols that enable the propagation and quantification of infectious virus. Here, we describe protocols for the propagation of SARS-CoV-2 in Vero E6 cells as well as two human cells lines, the intestinal epithelial Caco-2 cell line and the respiratory epithelial Calu-3 cell line. Additionally, we provide protocols for the quantification of SARS-CoV-2 by plaque assays and immunofocus forming assays in Vero E6 cells utilizing liquid overlays. These protocols provide a foundation for laboratories acquiring the ability to study SARS-CoV-2 to address this ongoing pandemic.
MeSH term(s)	Animals ; COVID-19 ; Caco-2 Cells ; Chlorocebus aethiops ; Humans ; Pandemics ; SARS-CoV-2 ; Vero Cells
Language	English
Publishing date	2022-05-12
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2111-0_8
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Structural Investigations of Interactions between the Influenza a Virus NS1 and Host Cellular Proteins.

Blake, Morgan E / Kleinpeter, Alex B / Jureka, Alexander S / Petit, Chad M

Viruses

2023 Volume 15, Issue 10

Abstract: The Influenza A virus is a continuous threat to public health that causes yearly epidemics with the ever-present threat of the virus becoming the next pandemic. Due to increasing levels of resistance, several of our previously used antivirals have been ... ...

Abstract	The Influenza A virus is a continuous threat to public health that causes yearly epidemics with the ever-present threat of the virus becoming the next pandemic. Due to increasing levels of resistance, several of our previously used antivirals have been rendered useless. There is a strong need for new antivirals that are less likely to be susceptible to mutations. One strategy to achieve this goal is structure-based drug development. By understanding the minute details of protein structure, we can develop antivirals that target the most conserved, crucial regions to yield the highest chances of long-lasting success. One promising IAV target is the virulence protein non-structural protein 1 (NS1). NS1 contributes to pathogenicity through interactions with numerous host proteins, and many of the resulting complexes have been shown to be crucial for virulence. In this review, we cover the NS1-host protein complexes that have been structurally characterized to date. By bringing these structures together in one place, we aim to highlight the strength of this field for drug discovery along with the gaps that remain to be filled.
MeSH term(s)	Humans ; Influenza A virus ; Immunity, Innate ; Virus Replication/genetics ; Interferons/metabolism ; Antiviral Agents/pharmacology ; Antiviral Agents/metabolism ; Viral Nonstructural Proteins/metabolism ; Influenza, Human ; Host-Pathogen Interactions/genetics
Chemical Substances	Interferons (9008-11-1) ; Antiviral Agents ; Viral Nonstructural Proteins
Language	English
Publishing date	2023-10-07
Publishing country	Switzerland
Document type	Journal Article ; Review ; Research Support, N.I.H., Extramural
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v15102063
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Pulsed Broad-Spectrum UV Light Effectively Inactivates SARS-CoV-2 on Multiple Surfaces and N95 Material.

Jureka, Alexander S / Williams, Caroline G / Basler, Christopher F

Viruses

2021 Volume 13, Issue 3

Abstract: The ongoing SARS-CoV-2 pandemic has resulted in an increased need for technologies capable of efficiently disinfecting public spaces as well as personal protective equipment. UV light disinfection is a well-established method for inactivating respiratory ...

Abstract	The ongoing SARS-CoV-2 pandemic has resulted in an increased need for technologies capable of efficiently disinfecting public spaces as well as personal protective equipment. UV light disinfection is a well-established method for inactivating respiratory viruses. Here, we have determined that broad-spectrum, pulsed UV light is effective at inactivating SARS-CoV-2 on multiple surfaces in vitro. For hard, non-porous surfaces, we observed that SARS-CoV-2 was inactivated to undetectable levels on plastic and glass with a UV dose of 34.9 mJ/cm
MeSH term(s)	COVID-19/prevention & control ; COVID-19/virology ; Disinfection/instrumentation ; Disinfection/methods ; Humans ; Masks/virology ; SARS-CoV-2/physiology ; SARS-CoV-2/radiation effects ; Ultraviolet Rays ; Virus Inactivation/radiation effects
Language	English
Publishing date	2021-03-11
Publishing country	Switzerland
Document type	Evaluation Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13030460
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Propagation, Inactivation, and Safety Testing of SARS-CoV-2.

Jureka, Alexander S / Silvas, Jesus A / Basler, Christopher F

Viruses

2020 Volume 12, Issue 6

Abstract: In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million ... ...

Abstract	In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly described and effective procedures that enable the propagation and quantification of infectious virus. As work with the virus is recommended to be performed at biosafety level 3, validated methods to effectively inactivate the virus to enable the safe study of RNA, DNA, and protein from infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat.
MeSH term(s)	Animals ; Betacoronavirus/drug effects ; Betacoronavirus/growth & development ; Betacoronavirus/pathogenicity ; Betacoronavirus/physiology ; COVID-19 ; Cellulose ; Chlorocebus aethiops ; Coronavirus Infections/virology ; Culture Media/chemistry ; Formaldehyde ; Guanidines/pharmacology ; HEK293 Cells ; Humans ; Pandemics ; Phenols/pharmacology ; Pneumonia, Viral/virology ; Propiolactone/pharmacology ; SARS-CoV-2 ; Sepharose ; Vero Cells ; Viral Plaque Assay/methods ; Virus Inactivation
Chemical Substances	Culture Media ; Guanidines ; Phenols ; trizol ; Formaldehyde (1HG84L3525) ; Propiolactone (6RC3ZT4HB0) ; Cellulose (9004-34-6) ; Sepharose (9012-36-6) ; microcrystalline cellulose (OP1R32D61U)
Keywords	covid19
Language	English
Publishing date	2020-06-06
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v12060622
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Pulsed broad-spectrum UV light effectively inactivates SARS-CoV-2 on multiple surfaces

Jureka, Alexander S / Williams, Caroline G / Basler, Christopher F

bioRxiv

Abstract	The ongoing SARS-CoV-2 pandemic has resulted in an increased need for technologies capable of efficiently disinfecting public spaces as well as personal protective equipment. UV light disinfection is a well-established method for inactivating respiratory viruses. Here, we have determined that broad-spectrum, pulsed UV light is effective at inactivating SARS-CoV-2 on multiple surfaces. For hard, non-porous surfaces we observed that SARS-CoV-2 was inactivated to undetectable levels on plastic and glass with a UV dose of 34.9 mJ/cm2 and stainless steel with a dose of 52.5 mJ/cm2. We also observed that broad-spectrum, pulsed UV light is effective at reducing SARS-CoV-2 on N95 respirator material to undetectable levels with a dose of 103 mJ/cm2. We included UV dosimeter cards that provide a colorimetric readout of UV dose and demonstrated their utility as a means to confirm desired levels of exposure were reached. Together, the results present here demonstrate that broad-spectrum, pulsed UV light is an effective technology for the inactivation of SARS-CoV-2 on multiple surfaces.
Keywords	covid19
Language	English
Publishing date	2021-02-15
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2021.02.12.431032
Database	COVID19

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Article: Pulsed Broad-Spectrum UV Light Effectively Inactivates SARS-CoV-2 on Multiple Surfaces and N95 Material

Jureka, Alexander S / Williams, Caroline G / Basler, Christopher F

Viruses. 2021 Mar. 11, v. 13, no. 3

2021

Abstract	The ongoing SARS-CoV-2 pandemic has resulted in an increased need for technologies capable of efficiently disinfecting public spaces as well as personal protective equipment. UV light disinfection is a well-established method for inactivating respiratory viruses. Here, we have determined that broad-spectrum, pulsed UV light is effective at inactivating SARS-CoV-2 on multiple surfaces in vitro. For hard, non-porous surfaces, we observed that SARS-CoV-2 was inactivated to undetectable levels on plastic and glass with a UV dose of 34.9 mJ/cm² and stainless steel with a dose of 52.5 mJ/cm². We also observed that broad-spectrum, pulsed UV light is effective at reducing SARS-CoV-2 on N95 respirator material to undetectable levels with a dose of 103 mJ/cm². We included UV dosimeter cards that provide a colorimetric readout of UV dose and demonstrated their utility as a means to confirm desired levels of exposure were reached. Together, the results presented here demonstrate that broad-spectrum, pulsed UV light is an effective technology for the in vitro inactivation of SARS-CoV-2 on multiple surfaces.
Keywords	Severe acute respiratory syndrome coronavirus 2 ; colorimetry ; disinfection ; glass ; pandemic ; safety equipment ; stainless steel ; ultraviolet radiation
Language	English
Dates of publication	2021-0311
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
Note	NAL-light
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13030460
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Propagation, inactivation, and safety testing of SARS-CoV-2

Jureka, Alexander S. / Silvas, Jesus A. / Basler, Christopher F.

bioRxiv

Abstract	In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly described and effective procedures that enable the propagation and quantification of infectious virus. Because work with the virus is recommended to be performed at biosafety level 3, validated methods to effectively inactivate the virus to enable safe study of RNA, DNA and protein from infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat.
Keywords	covid19
Publisher	BioRxiv; WHO
Document type	Article ; Online
DOI	10.1101/2020.05.13.094482
Database	COVID19

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Article ; Online: Propagation, inactivation, and safety testing of SARS-CoV-2

Jureka, Alexander S. / Silvas, Jesus A. / Basler, Christopher F.

bioRxiv

Abstract	In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly described and effective procedures that enable the propagation and quantification of infectious virus. Because work with the virus is recommended to be performed at biosafety level 3, validated methods to effectively inactivate the virus to enable safe study of RNA, DNA and protein from infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat.
Keywords	covid19
Language	English
Publishing date	2020-05-14
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2020.05.13.094482
Database	COVID19

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Article: Propagation, Inactivation, and Safety Testing of SARS-CoV-2

Jureka, Alexander S / Silvas, Jesus A / Basler, Christopher F

Viruses. 2020 June 06, v. 12, no. 6

2020

Abstract	In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly described and effective procedures that enable the propagation and quantification of infectious virus. As work with the virus is recommended to be performed at biosafety level 3, validated methods to effectively inactivate the virus to enable the safe study of RNA, DNA, and protein from infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat.
Keywords	COVID-19 infection ; DNA ; RNA ; Severe acute respiratory syndrome coronavirus 2 ; agarose ; biosafety ; cell lines ; cellulose ; formalin ; heat ; lactones ; pandemic ; pathogenicity ; safety testing ; viruses ; China
Language	English
Dates of publication	2020-0606
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v12060622
Database	NAL-Catalogue (AGRICOLA)

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Article: Inhibitors of VPS34 and lipid metabolism suppress SARS-CoV-2 replication.

Silvas, Jesus A / Jureka, Alexander S / Nicolini, Anthony M / Chvatal, Stacie A / Basler, Christopher F

bioRxiv : the preprint server for biology

2020

Abstract: Therapeutics targeting replication of SARS coronavirus 2 (SARS-CoV-2) are urgently needed. Coronaviruses rely on host membranes for entry, establishment of replication centers and egress. Compounds targeting cellular membrane biology and lipid ... ...

Abstract	Therapeutics targeting replication of SARS coronavirus 2 (SARS-CoV-2) are urgently needed. Coronaviruses rely on host membranes for entry, establishment of replication centers and egress. Compounds targeting cellular membrane biology and lipid biosynthetic pathways have previously shown promise as antivirals and are actively being pursued as treatments for other conditions. Here, we tested small molecule inhibitors that target membrane dynamics or lipid metabolism. Included were inhibitors of the PI3 kinase VPS34, which functions in autophagy, endocytosis and other processes; Orlistat, an inhibitor of lipases and fatty acid synthetase, is approved by the FDA as a treatment for obesity; and Triacsin C which inhibits long chain fatty acyl-CoA synthetases. VPS34 inhibitors, Orlistat and Triacsin C inhibited virus growth in Vero E6 cells and in the human airway epithelial cell line Calu-3, acting at a post-entry step in the virus replication cycle. Of these the VPS34 inhibitors exhibit the most potent activity.
Keywords	covid19
Language	English
Publishing date	2020-07-20
Publishing country	United States
Document type	Preprint
DOI	10.1101/2020.07.18.210211
Database	MEDical Literature Analysis and Retrieval System OnLINE

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