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Article ; Online: Contribution to pathogenesis of accessory proteins of deadly human coronaviruses.

Hurtado-Tamayo, Jesus / Requena-Platek, Ricardo / Enjuanes, Luis / Bello-Perez, Melissa / Sola, Isabel

Frontiers in cellular and infection microbiology

2023 Volume 13, Page(s) 1166839

Abstract: Coronaviruses (CoVs) are enveloped and positive-stranded RNA viruses with a large genome (∼ 30kb). CoVs include essential genes, such as the replicase and four genes coding for structural proteins (S, M, N and E), and genes encoding accessory proteins, ... ...

Abstract	Coronaviruses (CoVs) are enveloped and positive-stranded RNA viruses with a large genome (∼ 30kb). CoVs include essential genes, such as the replicase and four genes coding for structural proteins (S, M, N and E), and genes encoding accessory proteins, which are variable in number, sequence and function among different CoVs. Accessory proteins are non-essential for virus replication, but are frequently involved in virus-host interactions associated with virulence. The scientific literature on CoV accessory proteins includes information analyzing the effect of deleting or mutating accessory genes in the context of viral infection, which requires the engineering of CoV genomes using reverse genetics systems. However, a considerable number of publications analyze gene function by overexpressing the protein in the absence of other viral proteins. This ectopic expression provides relevant information, although does not acknowledge the complex interplay of proteins during virus infection. A critical review of the literature may be helpful to interpret apparent discrepancies in the conclusions obtained by different experimental approaches. This review summarizes the current knowledge on human CoV accessory proteins, with an emphasis on their contribution to virus-host interactions and pathogenesis. This knowledge may help the search for antiviral drugs and vaccine development, still needed for some highly pathogenic human CoVs.
MeSH term(s)	Humans ; Coronavirus/genetics ; Coronavirus Infections ; Viral Proteins/genetics ; Antiviral Agents ; Virulence
Chemical Substances	Viral Proteins ; Antiviral Agents
Language	English
Publishing date	2023-05-01
Publishing country	Switzerland
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	2619676-1
ISSN	2235-2988 ; 2235-2988
ISSN (online)	2235-2988
ISSN	2235-2988
DOI	10.3389/fcimb.2023.1166839
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Viricidal Activity of Thermoplastic Polyurethane Materials with Silver Nanoparticles.

Díaz-Puertas, Rocío / Rodríguez-Cañas, Enrique / Bello-Perez, Melissa / Fernández-Oliver, Marta / Mallavia, Ricardo / Falco, Alberto

Nanomaterials (Basel, Switzerland)

2023 Volume 13, Issue 9

Abstract: The use of diverse Ag-based nanoparticulated forms has shown promising results in controlling viral propagation. In this study, a commercial nanomaterial consisting of ceramic-coated silver nanoparticles (AgNPs) was incorporated into thermoplastic ... ...

Abstract	The use of diverse Ag-based nanoparticulated forms has shown promising results in controlling viral propagation. In this study, a commercial nanomaterial consisting of ceramic-coated silver nanoparticles (AgNPs) was incorporated into thermoplastic polyurethane (TPU) plates using an industrial protocol, and the surface composition, ion-release dynamics and viricidal properties were studied. The surface characterization by FESEM-EDX revealed that the molar composition of the ceramic material was 5.5 P:3.3 Mg:Al and facilitated the identification of the embedded AgNPs (54.4 ± 24.9 nm). As determined by ICPMS, the release rates from the AgNP-TPU into aqueous solvents were 4 ppm/h for Ag and Al, and 28.4 ppm/h for Mg ions. Regarding the biological assays, the AgNP-TPU material did not induce significant cytotoxicity in the cell lines employed. Its viricidal activity was characterized, based on ISO 21702:2019, using the Spring viraemia of carp virus (SVCV), and then tested against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The results demonstrated that AgNP-TPU materials exhibited significant (75%) and direct antiviral activity against SVCV virions in a time- and temperature-dependent manner. Similar inhibition levels were found against SARS-CoV-2. These findings show the potential of AgNP-TPU-based materials as a supporting strategy to control viral spread.
Language	English
Publishing date	2023-04-25
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662255-5
ISSN	2079-4991
ISSN	2079-4991
DOI	10.3390/nano13091467
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Article: Update on the Inactivation Procedures for the Vaccine Development Prospects of a New Highly Virulent RGNNV Isolate.

Falco, Alberto / Bello-Perez, Melissa / Díaz-Puertas, Rocío / Mold, Matthew / Adamek, Mikolaj

Vaccines

2021 Volume 9, Issue 12

Abstract: Viral nervous necrosis (VNN) caused by the nervous necrosis virus (NNV) affects a broad range of primarily marine fish species, with mass mortality rates often seen among larvae and juveniles. Its genetic diversification may hinder the effective ... ...

Abstract	Viral nervous necrosis (VNN) caused by the nervous necrosis virus (NNV) affects a broad range of primarily marine fish species, with mass mortality rates often seen among larvae and juveniles. Its genetic diversification may hinder the effective implementation of preventive measures such as vaccines. The present study describes different inactivation procedures for developing an inactivated vaccine against a new NNV isolate confirmed to possess deadly effects upon the European seabass (
Language	English
Publishing date	2021-12-07
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines9121441
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Article: Modulation of the Tissue Expression Pattern of Zebrafish CRP-Like Molecules Suggests a Relevant Antiviral Role in Fish Skin

Bello-Perez, Melissa / Adamek, Mikolaj / Coll, Julio / Figueras, Antonio / Novoa, Beatriz / Falco, Alberto

Biology. 2021 Jan. 22, v. 10, no. 2

2021

Abstract: Recent studies suggest that short pentraxins in fish might serve as biomarkers for not only bacterial infections, as in higher vertebrates including humans, but also for viral ones. These fish orthologs of mammalian short pentraxins are currently ... ...

Abstract	Recent studies suggest that short pentraxins in fish might serve as biomarkers for not only bacterial infections, as in higher vertebrates including humans, but also for viral ones. These fish orthologs of mammalian short pentraxins are currently attracting interest because of their newly discovered antiviral activity. In the present work, the modulation of the gene expression of all zebrafish short pentraxins (CRP-like proteins, CRP1-7) was extensively analyzed by quantitative polymerase chain reaction. Initially, the tissue distribution of crp1-7 transcripts and how the transcripts varied in response to a bath infection with the spring viremia of carp virus, were determined. The expression of crp1-7 was widely distributed and generally increased after infection (mostly at 5 days post infection), except for crp1 (downregulated). Interestingly, several crp transcription levels significantly increased in skin. Further assays in mutant zebrafish of recombinant activation gene 1 (rag1) showed that all crps (except for crp2, downregulated) were already constitutively highly expressed in skin from rag1 knockouts and only increased moderately after viral infection. Similar results were obtained for most mx isoforms (a reporter gene of the interferon response), suggesting a general overcompensation of the innate immunity in the absence of the adaptive one.
Keywords	Carp sprivivirus ; Danio rerio ; antiviral properties ; biomarkers ; fish ; fish skin ; gene expression ; innate immunity ; interferons ; mutants ; quantitative polymerase chain reaction ; reporter genes ; tissue distribution
Language	English
Dates of publication	2021-0122
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	2661517-4
ISSN	2079-7737
ISSN	2079-7737
DOI	10.3390/biology10020078
Database	NAL-Catalogue (AGRICOLA)

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Article: Modulation of the Tissue Expression Pattern of Zebrafish CRP-Like Molecules Suggests a Relevant Antiviral Role in Fish Skin.

Bello-Perez, Melissa / Adamek, Mikolaj / Coll, Julio / Figueras, Antonio / Novoa, Beatriz / Falco, Alberto

Biology

2021 Volume 10, Issue 2

Abstract	Recent studies suggest that short pentraxins in fish might serve as biomarkers for not only bacterial infections, as in higher vertebrates including humans, but also for viral ones. These fish orthologs of mammalian short pentraxins are currently attracting interest because of their newly discovered antiviral activity. In the present work, the modulation of the gene expression of all zebrafish short pentraxins (CRP-like proteins, CRP1-7) was extensively analyzed by quantitative polymerase chain reaction. Initially, the tissue distribution of
Language	English
Publishing date	2021-01-22
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2661517-4
ISSN	2079-7737
ISSN	2079-7737
DOI	10.3390/biology10020078
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Article ; Online: Viricidal Activity of Thermoplastic Polyurethane Materials with Silver Nanoparticles

Rocío Díaz-Puertas / Enrique Rodríguez-Cañas / Melissa Bello-Perez / Marta Fernández-Oliver / Ricardo Mallavia / Alberto Falco

Nanomaterials, Vol 13, Iss 1467, p

2023 Volume 1467

Abstract	The use of diverse Ag-based nanoparticulated forms has shown promising results in controlling viral propagation. In this study, a commercial nanomaterial consisting of ceramic-coated silver nanoparticles (AgNPs) was incorporated into thermoplastic polyurethane (TPU) plates using an industrial protocol, and the surface composition, ion-release dynamics and viricidal properties were studied. The surface characterization by FESEM-EDX revealed that the molar composition of the ceramic material was 5.5 P:3.3 Mg:Al and facilitated the identification of the embedded AgNPs (54.4 ± 24.9 nm). As determined by ICPMS, the release rates from the AgNP–TPU into aqueous solvents were 4 ppm/h for Ag and Al, and 28.4 ppm/h for Mg ions. Regarding the biological assays, the AgNP–TPU material did not induce significant cytotoxicity in the cell lines employed. Its viricidal activity was characterized, based on ISO 21702:2019, using the Spring viraemia of carp virus (SVCV), and then tested against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The results demonstrated that AgNP–TPU materials exhibited significant (75%) and direct antiviral activity against SVCV virions in a time- and temperature-dependent manner. Similar inhibition levels were found against SARS-CoV-2. These findings show the potential of AgNP–TPU-based materials as a supporting strategy to control viral spread.
Keywords	3D nanomaterials ; thermoplastic polyurethane ; ceramic ; Ag nanoparticles ; viricidal activity ; Chemistry ; QD1-999
Subject code	660
Language	English
Publishing date	2023-04-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Zebrafish C-reactive protein isoforms inhibit SVCV replication by blocking autophagy through interactions with cell membrane cholesterol.

Bello-Perez, Melissa / Pereiro, Patricia / Coll, Julio / Novoa, Beatriz / Perez, Luis / Falco, Alberto

Scientific reports

2020 Volume 10, Issue 1, Page(s) 566

Abstract: In the present work, the mechanisms involved in the recently reported antiviral activity of zebrafish C-reactive protein-like protein (CRP1-7) against the spring viraemia of carp rhabdovirus (SVCV) in fish are explored. The results neither indicate ... ...

Abstract	In the present work, the mechanisms involved in the recently reported antiviral activity of zebrafish C-reactive protein-like protein (CRP1-7) against the spring viraemia of carp rhabdovirus (SVCV) in fish are explored. The results neither indicate blocking of the attachment or the binding step of the viral replication cycle nor suggest the direct inhibition of G protein fusion activity or the stimulation of the host's interferon system. However, an antiviral state in the host is induced. Further results showed that the antiviral protection conferred by CRP1-7 was mainly due to the inhibition of autophagic processes. Thus, given the high affinity of CRPs for cholesterol and the recently described influence of the cholesterol balance in lipid rafts on autophagy, both methyl-β-cyclodextrin (a cholesterol-complexing agent) and 25-hydroxycholesterol (a cholesterol molecule with antiviral properties) were used to further describe CRP activity. All the tested compounds exerted antiviral activity by affecting autophagy in a similar manner. Further assays indicate that CRP reduces autophagy activity by initially disturbing the cholesterol ratios in the host cellular membranes, which in turn negatively affects the intracellular regulation of reactive oxygen species (ROS) and increases lysosomal pH as a consequence. Ultimately, here we propose that such pH changes exert an inhibitory direct effect on SVCV replication by disrupting the pH-dependent membrane-fusogenic ability of the viral glycoprotein G, which allows the release of the virus from endosomes into cytoplasm during its entry phase.
MeSH term(s)	Animals ; Autophagy ; C-Reactive Protein/genetics ; C-Reactive Protein/pharmacology ; Cell Line ; Cell Membrane/chemistry ; Cholesterol/metabolism ; Hydrogen-Ion Concentration/drug effects ; Hydroxycholesterols/metabolism ; Protein Isoforms/pharmacology ; Reactive Oxygen Species/metabolism ; Rhabdoviridae/drug effects ; Rhabdoviridae/physiology ; Rhabdoviridae Infections/metabolism ; Rhabdoviridae Infections/prevention & control ; Virus Replication/drug effects ; Zebrafish/genetics ; Zebrafish/virology ; Zebrafish Proteins/genetics ; Zebrafish Proteins/pharmacology ; beta-Cyclodextrins/metabolism
Chemical Substances	Hydroxycholesterols ; Protein Isoforms ; Reactive Oxygen Species ; Zebrafish Proteins ; beta-Cyclodextrins ; methyl-beta-cyclodextrin ; 25-hydroxycholesterol (767JTD2N31) ; C-Reactive Protein (9007-41-4) ; Cholesterol (97C5T2UQ7J)
Language	English
Publishing date	2020-01-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-020-57501-0
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Discovery of nonnucleoside inhibitors of polymerase from infectious pancreatic necrosis virus (IPNV).

Bello-Pérez, Melissa / Falcó, Alberto / Galiano, Vicente / Coll, Julio / Perez, Luis / Encinar, José Antonio

Drug design, development and therapy

2018 Volume 12, Page(s) 2337–2359

Abstract: Introduction: Infectious pancreatic necrosis virus (IPNV) causes serious losses in several fish species of commercial interest. IPNV is a non-enveloped double-stranded RNA virus with a genome consisting of two segments A and B. Segment B codes for the ... ...

Abstract	Introduction: Infectious pancreatic necrosis virus (IPNV) causes serious losses in several fish species of commercial interest. IPNV is a non-enveloped double-stranded RNA virus with a genome consisting of two segments A and B. Segment B codes for the VP1 protein, a non-canonical RNA-dependent RNA polymerase that can be found both in its free form and linked to the end of genomic RNA, an essential enzyme for IPNV replication. Materials and methods: We take advantage of the knowledge over the allosteric binding site described on the surface of the thumb domain of Hepatitis C virus (HCV) polymerase to design new non-nucleoside inhibitors against the IPNV VP1 polymerase. Results: Molecular docking techniques have been used to screen a chemical library of 23,760 compounds over a defined cavity in the surface of the thumb domain. Additional ADMET (absorption, distribution, metabolism, excretion, and toxicity) filter criteria has been applied. Conclusion: We select two sets of 9 and 50 inhibitor candidates against the polymerases of HCV and IPNV, respectively. Two non-toxic compounds have been tested in vitro with antiviral capacity against IPNV Sp and LWVRT60 strains in the low µM range with different activity depending on the IPNV strain used.
MeSH term(s)	Antiviral Agents/pharmacology ; Cell Survival/drug effects ; Cells, Cultured ; Drug Discovery ; Hepacivirus/drug effects ; Infectious pancreatic necrosis virus/drug effects ; Infectious pancreatic necrosis virus/enzymology ; Molecular Docking Simulation ; RNA-Dependent RNA Polymerase/antagonists & inhibitors ; RNA-Dependent RNA Polymerase/chemistry
Chemical Substances	Antiviral Agents ; RNA-Dependent RNA Polymerase (EC 2.7.7.48)
Language	English
Publishing date	2018-07-30
Publishing country	New Zealand
Document type	Journal Article
ZDB-ID	2451346-5
ISSN	1177-8881 ; 1177-8881
ISSN (online)	1177-8881
ISSN	1177-8881
DOI	10.2147/DDDT.S171087
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Article ; Online: Canonical and Noncanonical Autophagy as Potential Targets for COVID-19.

Bello-Perez, Melissa / Sola, Isabel / Novoa, Beatriz / Klionsky, Daniel J / Falco, Alberto

Cells

2020 Volume 9, Issue 7

Abstract: The SARS-CoV-2 pandemic necessitates a review of the molecular mechanisms underlying cellular infection by coronaviruses, in order to identify potential therapeutic targets against the associated new disease (COVID-19). Previous studies on its ... ...

Abstract	The SARS-CoV-2 pandemic necessitates a review of the molecular mechanisms underlying cellular infection by coronaviruses, in order to identify potential therapeutic targets against the associated new disease (COVID-19). Previous studies on its counterparts prove a complex and concomitant interaction between coronaviruses and autophagy. The precise manipulation of this pathway allows these viruses to exploit the autophagy molecular machinery while avoiding its protective apoptotic drift and cellular innate immune responses. In turn, the maneuverability margins of such hijacking appear to be so narrow that the modulation of the autophagy, regardless of whether using inducers or inhibitors (many of which are FDA-approved for the treatment of other diseases), is usually detrimental to viral replication, including SARS-CoV-2. Recent discoveries indicate that these interactions stretch into the still poorly explored noncanonical autophagy pathway, which might play a substantial role in coronavirus replication. Still, some potential therapeutic targets within this pathway, such as RAB9 and its interacting proteins, look promising considering current knowledge. Thus, the combinatory treatment of COVID-19 with drugs affecting both canonical and noncanonical autophagy pathways may be a turning point in the fight against this and other viral infections, which may also imply beneficial prospects of long-term protection.
MeSH term(s)	Antiviral Agents/pharmacology ; Antiviral Agents/therapeutic use ; Apoptosis ; Autophagy/drug effects ; Autophagy-Related Proteins/antagonists & inhibitors ; Autophagy-Related Proteins/metabolism ; Betacoronavirus/classification ; Betacoronavirus/physiology ; COVID-19 ; Capsid Proteins/metabolism ; Coronavirus Infections/drug therapy ; Coronavirus Infections/pathology ; Coronavirus Infections/virology ; Coronavirus Nucleocapsid Proteins ; Humans ; Pandemics ; Pneumonia, Viral/drug therapy ; Pneumonia, Viral/pathology ; Pneumonia, Viral/virology ; SARS-CoV-2 ; Virus Replication/drug effects
Chemical Substances	Antiviral Agents ; Autophagy-Related Proteins ; Capsid Proteins ; Coronavirus Nucleocapsid Proteins ; NSP6 protein, SARS-CoV-2
Keywords	covid19
Language	English
Publishing date	2020-07-05
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells9071619
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Article ; Online: Update on the Inactivation Procedures for the Vaccine Development Prospects of a New Highly Virulent RGNNV Isolate

Alberto Falco / Melissa Bello-Perez / Rocío Díaz-Puertas / Matthew Mold / Mikolaj Adamek

Vaccines, Vol 9, Iss 1441, p

2021 Volume 1441

Abstract	Viral nervous necrosis (VNN) caused by the nervous necrosis virus (NNV) affects a broad range of primarily marine fish species, with mass mortality rates often seen among larvae and juveniles. Its genetic diversification may hinder the effective implementation of preventive measures such as vaccines. The present study describes different inactivation procedures for developing an inactivated vaccine against a new NNV isolate confirmed to possess deadly effects upon the European seabass ( Dicentrarchus labrax ), an important Mediterranean farmed fish species that is highly susceptible to this disease. First, an NNV isolate from seabass adults diagnosed with VNN was rescued and the sequences of its two genome segments (RNA1 and RNA2) were classified into the red-spotted grouper NNV (RGNNV) genotype, closely clustering to the highly pathogenic 283.2009 isolate. The testing of different inactivation procedures revealed that the virus particles of this isolate showed a marked resistance to heat (for at least 60 °C for 120 min with and without 1% BSA) but that they were fully inactivated by 3 mJ/cm 2 UV-C irradiation and 24 h 0.2% formalin treatment, which stood out as promising NNV-inactivation procedures for potential vaccine candidates. Therefore, these procedures are feasible, effective, and rapid response strategies for VNN control in aquaculture.
Keywords	viral nervous necrosis ; betanodavirus ; nervous necrosis virus ; autogenous vaccine ; virus inactivation ; virus-inactivated vaccine ; Medicine ; R
Subject code	590
Language	English
Publishing date	2021-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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