The Antiviral Molecule 5-Pyridoxolactone Identified Post BmNPV Infection of the Silkworm, Bombyx mori

Bombyx mori nucleopolyhedrovirus (BmNPV) is a pathogen that causes great economic losses in sericulture. Many genes play a role in viral infection of silkworms, but silkworm metabolism in response to BmNPV infection is unknown. We studied BmE cells infected with BmNPV. We performed liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS)-based non-targeted metabolomics analysis of the cytosolic extract and identified 36, 76, 138, 101, 189, and 166 different molecules at 3, 6, 12, 24, 48, and 72 h post BmNPV infection (hpi) compared with 0 hpi. Compounds representing different areas of metabolism were increased in cells post BmNPV infection. These areas included purine metabolism, aminoacyl−tRNA biosynthesis, and ABC transporters. Glycerophosphocholine (GPC), 2-hydroxyadenine (2-OH-Ade), gamma-glutamylcysteine (γ-Glu-Cys), hydroxytolbutamide, and 5-pyridoxolactone glycerophosphocholine were continuously upregulated in BmE cells post BmNPV infection by heat map analysis. Only 5-pyridoxolactone was found to strongly inhibit the proliferation of BmNPV when it was used to treat BmE cells. Fewer infected cells were detected and the level of BmNPV DNA decreased with increasing 5-pyridoxolactone in a dose-dependent manner. The expression of BmNPV genes ie1, helicase, GP64, and VP39 in BmE cells treated with 5-pyridoxolactone were strongly inhibited in the BmNPV infection stage. This suggested that 5-pyridoxolactone may suppress the entry of BmNPV. The data in this study characterize the metabolism changes in BmNPV-infected cells. Further analysis of 5-pyridoxolactone, which is a robust antiviral molecule, may increase our understanding of antiviral immunity.

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Abstract P492: Putative Protein Biomarkers For Reductive Stress Cardiomyopathy

Circulation Research ◽

10.1161/res.129.suppl_1.p492 ◽

2021 ◽

Vol 129 (Suppl_1) ◽

Author(s):

Rajasekaran Namakkal-Soorappan ◽

Cynthia L David ◽

Krishna Parsawar

Keyword(s):

Protein Quality ◽

Redox Homeostasis ◽

Differentially Expressed ◽

Related Factor ◽

Protein Biomarkers ◽

Heat Map ◽

Reductive Stress ◽

Tandem Mass Tag ◽

Map Analysis ◽

Dose Dependent

Background: Nuclear factor erythroid 2-related factor 2 (NRF2) signaling is vital for redox homeostasis. We reported that transgenic mice expressing constitutively active Nrf2 (CaNrf2) exhibit reductive stress (RS). Here in, we identified novel protein signatures reacting to RS-induced cardiomyopathy. Methods: Tandem Mass Tag (TMT) proteomic analysis was performed in the heart tissues of Ca-Nrf2-transgenic (TG-low & TG-high) and non-transgenic (NTg) mice at 6 months of age (N= 4/group). Differentially expressed proteins (DEPs) were then identified using Scaffold. Validated the key DEPs using immunoblotting. PANTHER and STRING analysis were used to identify potential targets and their interactions. Results: A total of 1105 proteins were extracted from 24369 spectra. Of note, 226 and 261 proteins were differentially expressed in TG-L and TG-H vs. NTg hearts indicating a unique proteome signature for RS. Heat map analysis revealed a clear distinction between the TG-L and TG-H due to the dose-dependent effects of transgene/RS. Majority of the DEPs that are significantly altered in RS mice found to involve in stress related pathways such as antioxidants, NADPH, protein quality control (PQC), etc. Interestingly, some of these proteins were redox modified at their cysteine residues under chronic RS setting. Conclusions: TMT based proteomic analyses revealed unique proteome signatures for RS. The cysteine modifications in multiple proteins likely to cause pathological alterations via impaired PQC mechanisms. Molecular studies related to RS-mediated redox modifications in structural and functional cardiac proteome are underway.

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Identification of Diphenoxylate as an Antiviral Agent Against Severe Acute Respiratory Syndrome Coronavirus 2

10.26434/chemrxiv.12496547.v1 ◽

2020 ◽

Author(s):

Jin Soo Shin ◽

Eunhye Jung ◽

Yejin Jang ◽

Soo Bong Han ◽

Meehyein Kim

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Antiviral Treatment ◽

Antiviral Agent ◽

Selectivity Index ◽

Dependent Manner ◽

S Protein ◽

Chemical Derivatives ◽

Highly Active ◽

Infected Cells ◽

Dose Dependent

This is the first report to show that diphenoxylate is highly active against SARS-CoV-2 with EC<sub>50</sub> of 1.4 μM, CC<sub>50</sub> over 100 μM and selectivity index over 71.4 after screening of 14 diphenyl derivatives. Our results with highly purified solid diphenoxylate confirmatively demonstrate that the viral S protein is reduced in the virus-infected cells in a dose-dependent manner. It could provide insights for antiviral treatment of COVID-19 with diphenoxylate or its chemical derivatives

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The novel antiviral properties of brassicasterol against human adenovirus

Future Virology ◽

10.2217/fvl-2021-0087 ◽

2021 ◽

Author(s):

Peifeng Yu ◽

Dan Lou ◽

Lifeng Qi ◽

Zewei Chen

Keyword(s):

Dna Replication ◽

Epithelial Cells ◽

Viral Entry ◽

Human Adenovirus ◽

Airway Epithelial Cells ◽

Dependent Manner ◽

The Novel ◽

Airway Epithelial ◽

Infected Cells ◽

Dose Dependent

Aim: To investigate whether brassicasterol has inhibitory effects against adenovirus (AdV). Materials and methods: The antiviral effects of brassicasterol against AdV 3 and 7 were tested in human airway epithelial cells. Brassicasterol cytotoxicity was assessed by WST-1 assay. AdV DNA was quantified by qPCR. Results: Brassicasterol inhibited AdV 3 and 7 infection of airway epithelial cells in a dose-dependent manner. Similarly, brassicasterol also inhibited AdV 3 and 7 production in infected cells. No apparent cytotoxicity of brassicasterol was detected. Further study showed that brassicasterol inhibited AdV DNA replication, but had no impact on viral entry of cells and viral genome import to nucleus. Conclusion: Brassicasterol exerts anti-AdV effects probably through the inhibition of viral DNA replication.

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Inhibition of Mycotoxigenic Fungi in Different Vegetable Matrices by Extracts of Trichoderma Species

Journal of Fungi ◽

10.3390/jof7060445 ◽

2021 ◽

Vol 7 (6) ◽

pp. 445

Author(s):

Claudia Stracquadanio ◽

Carlos Luz ◽

Federico La Spada ◽

Giuseppe Meca ◽

Santa Olga Cacciola

Keyword(s):

Fusarium Verticillioides ◽

Economic Losses ◽

Dependent Manner ◽

Trichoderma Atroviride ◽

Penicillium Verrucosum ◽

Time Intervals ◽

Trichoderma Species ◽

Mycotoxigenic Fungi ◽

Dose Dependent ◽

Chemical Fungicides

Post-harvest fungal diseases of plant products are a serious concern leading to economic losses and health risks. Moreover, the use of synthetic chemical fungicides to prevent these diseases is limited due to toxic residues. This study aimed at determining the effective dose of extracts of Trichoderma asperellum IMI393899 (TE1) and Trichoderma atroviride TS (TE2) in inhibiting the contamination by mycotoxigenic fungi on different plant matrices. Extracts were tested on tomatoes contaminated by Fusarium verticillioides and Fusarium graminearum, wheat contaminated by Penicillium verrucosum and maize contaminated by Aspergillus flavus. The efficacy of extracts was evaluated at two time intervals after treatment, 4 and 11 days for tomato, and 10 and 20 days for both wheat and maize. Both extracts showed a significant inhibitory activity on mycotoxigenic pathogens and significantly reduced Log CFU/g compared to the control. Moreover, the extracts reduced mycotoxin production in a dose dependent manner and with a long-lasting effect. The ochratoxin A was reduced by both extracts but only the extract TE2 was effective in reducing aflatoxins, whereas TE1 treatment increased their synthesis.

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Bepridil is potent against SARS-CoV-2 in vitro

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2012201118 ◽

2021 ◽

Vol 118 (10) ◽

pp. e2012201118 ◽

Cited By ~ 1

Author(s):

Erol C. Vatansever ◽

Kai S. Yang ◽

Aleksandra K. Drelich ◽

Kaci C. Kratch ◽

Chia-Chuan Cho ◽

...

Keyword(s):

European Medicines Agency ◽

Dependent Manner ◽

Live Virus ◽

Docking Analysis ◽

Computational Docking ◽

Main Protease ◽

Infected Cells ◽

Endosomal Ph ◽

Dose Dependent

Guided by a computational docking analysis, about 30 Food and Drug Administration/European Medicines Agency (FDA/EMA)-approved small-molecule medicines were characterized on their inhibition of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro). Of these small molecules tested, six displayed a concentration that inhibits response by 50% (IC50) value below 100 μM in inhibiting Mpro, and, importantly, three, that is, pimozide, ebastine, and bepridil, are basic molecules that potentiate dual functions by both raising endosomal pH to interfere with SARS-CoV-2 entry into the human cell host and inhibiting Mpro in infected cells. A live virus-based modified microneutralization assay revealed that bepridil possesses significant anti−SARS-CoV-2 activity in both Vero E6 and A459/ACE2 cells in a dose-dependent manner with low micromolar effective concentration, 50% (EC50) values. Therefore, the current study urges serious considerations of using bepridil in COVID-19 clinical tests.

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Identification of Diphenoxylate as an Antiviral Agent Against Severe Acute Respiratory Syndrome Coronavirus 2

10.26434/chemrxiv.12496547 ◽

2020 ◽

Author(s):

Jin Soo Shin ◽

Eunhye Jung ◽

Yejin Jang ◽

Soo Bong Han ◽

Meehyein Kim

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Antiviral Treatment ◽

Antiviral Agent ◽

Selectivity Index ◽

Dependent Manner ◽

S Protein ◽

Chemical Derivatives ◽

Highly Active ◽

Infected Cells ◽

Dose Dependent

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Bombyx mori Nucleopolyhedrovirus p26 Is Associated with Viral Late Stage Replication

Insects ◽

10.3390/insects12080707 ◽

2021 ◽

Vol 12 (8) ◽

pp. 707

Author(s):

Jun-Qing Ge ◽

Zhu-Hong Wang ◽

Xi Chen ◽

Hua Chen ◽

Jian Huang

Keyword(s):

Infected Cell ◽

Bombyx Mori ◽

Infectious Virus ◽

Pcr Analysis ◽

Immunofluorescence Analysis ◽

Bombyx Mori Nucleopolyhedrovirus ◽

Cell Cytoplasm ◽

Infected Cells ◽

Budded Virus ◽

Infection Stage

Bombyx mori nucleopolyhedrovirus (BmNPV) p26 is conserved among all Lepidoptera baculoviruses that have been completely sequenced thus far, and some baculoviruses even have two copies of p26, which suggested that p26 may play an important role in the virus infection cycle. This study aimed to characterize BmNPV p26. We found that BmNPV p26 transcripts were detectable as early as 3 h post-infection (hpi), and the transcript levels rapidly increased starting from 12 hpi. Western blot analysis using an anti-p26 polyclonal antibody demonstrated that the corresponding protein was also detectable from 6 hpi in BmNPV-infected cell lysates. Immunofluorescence analysis demonstrated that p26 was mainly dispersed in the infected cell cytoplasm, whereas the over-expressed fusion protein EGFP-p26 also accumulated in the nucleus. These results indicated that p26 is an early BmNPV gene and has functions both in the cytoplasm and the nucleus. RNAi-based knockdown of p26 could produce infectious virus and normal-appearing virions but decreased budded virus (BV) production in BmNPV-infected cells at 72 hpi. Moreover, the results of further quantitative PCR (Q-PCR) analysis indicated that the gp64 and p74 transcripts levels decreased significantly. These results indicated that BmNPV p26 may be associated with BmNPV replication during the late infection stage.

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Antiviral Activity of Germacrone against Pseudorabies Virus in Vitro

Pathogens ◽

10.3390/pathogens8040258 ◽

2019 ◽

Vol 8 (4) ◽

pp. 258 ◽

Cited By ~ 1

Author(s):

Wanting He ◽

Xiaofeng Zhai ◽

Jingyin Su ◽

Rui Ye ◽

Yuna Zheng ◽

...

Keyword(s):

Antiviral Activity ◽

Pseudorabies Virus ◽

Receptor Protein ◽

Economic Losses ◽

Dependent Manner ◽

Treatment And Prevention ◽

Novel Variant ◽

Acute Infectious Disease ◽

Dose Dependent

Pseudorabies virus (PRV), a member of the Herpesviridae, is the causative agent of an acute infectious disease in a variety of animals. The emergence of a novel variant strain brought huge economic losses to the pig industry since classical vaccine strains were not completely effective against variant strains. Therefore, the development of new anti-pseudorabies virus drugs and vaccines is of great significance for the treatment and prevention of pseudorabies. In this study, we found that germacrone, one of the major components of the essential oils extracted from Rhizoma Curcuma, was able to effectively inhibit PRV replication in a dose-dependent manner in vitro. Germacrone showed antiviral activity against PRV in the early phase of the viral replication cycle. Moreover, we found that germacrone does not directly kill the virus, nor does it affect the expression of the PRV receptor protein nectin-1, nectin-2, and CD155. Our results suggest germacrone could be used as an efficient microbicide or immunomodulatory agent in the control of the emerging variant PRV.

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Metabolism and Selective Toxicity of 6-Nitrobenzylthioinosine in Toxoplasma gondii

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.43.10.2437 ◽

1999 ◽

Vol 43 (10) ◽

pp. 2437-2443 ◽

Cited By ~ 27

Author(s):

Mahmoud H. el Kouni ◽

Vincenzo Guarcello ◽

Omar N. Al Safarjalani ◽

Fardos N. M. Naguib

Keyword(s):

Toxoplasma Gondii ◽

Human Fibroblasts ◽

Adenosine Kinase ◽

Host Cells ◽

Dependent Manner ◽

Selective Toxicity ◽

Link Type ◽

Infected Cells ◽

Dose Dependent ◽

Uninfected Host

ABSTRACT The purine nucleoside analogue NBMPR {nitrobenzylthioinosine or 6-[(4-nitrobenzyl)thio]-9-β-d-ribofuranosylpurine} was selectively phosphorylated to its nucleoside 5′-monophosphate byToxoplasma gondii but not mammalian adenosine kinase (EC2.7.1.20). NBMPR was also cleaved in toxoplasma to its nucleobase, nitrobenzylmercaptopurine. However, nitrobenzylmercaptopurine was not a substrate for either adenosine kinase or hypoxanthine-guanine-xanthine phosphoribosyltransferase (EC 2.4.2.8). Because of this unique and previously unknown metabolism of NBMPR by the parasite, the effect of NBMPR as an antitoxoplasmic agent was tested. NBMPR killed T. gondii grown in human fibroblasts in a dose-dependent manner, with a 50% inhibitory concentration of approximately 10 μM and without apparent toxicity to host cells. Doses of up to 100 μM had no significant toxic effect on uninfected host cells. The promising antitoxoplasmic effect of NBMPR led to the testing of other 6-substituted 9-β-d-ribofuranosylpurines, which were shown to be good ligands of the parasite adenosine kinase (M. H. Iltzsch, S. S. Uber, K. O. Tankersley, and M. H. el Kouni, Biochem. Pharmacol. 49:1501–1512, 1995), as antitoxoplasmic agents. Among the analogues tested, 6-benzylthioinosine,p-nitrobenzyl-6-selenopurine riboside,N 6-(p-azidobenzyl)adenosine, andN 6-(p-nitrobenzyl)adenosine, like NBMPR, were selectively toxic to parasite-infected cells. Thus, it appears that the unique characteristics of purine metabolism inT. gondii render certain 6-substituted 9-β-d-ribofuranosylpurines promising antitoxoplasmic drugs.

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Antiviral Activities of Mulberry (Morus alba) Juice and Seed against Influenza Viruses

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2018/2606583 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10 ◽

Cited By ~ 3

Author(s):

Hyojin Kim ◽

Mi Sook Chung

Keyword(s):

Radical Scavenging ◽

Antiviral Effect ◽

Morus Alba ◽

Influenza Viruses ◽

Dependent Manner ◽

Antiviral Activities ◽

2009 H1n1 ◽

Infected Cells ◽

A Minor ◽

Dose Dependent

Antiviral activities of Morus alba (MA) juice and seed were examined using time-of-addition plaque assays against influenza viruses, A/Brisbane/59/2007 (H1N1) (BR59), pandemic A/Korea/01/2009(H1N1) (KR01), A/Brisbane/10/2007(H3N2) (BR10), and B/Florida/4/2006 (FL04). MA juice (MAJ) showed much higher antiviral activity than MA seed (MAS). In the pre- and cotreatment of virus, MAJ showed antiviral effects against BR59, KR01, and FL04 in a dose-dependent manner. In particular, MAJ at 4% concentration exhibited 1.3 log inhibition in the pre- and cotreatment of the virus against FL04, a type B virus. However, little or weak inhibition was observed in the posttreatment of MAJ. GSH levels in the virus-infected cells were also examined. The decreased levels by the viral infection were restored significantly by the addition of MAJ. MAJ also exhibited significant DPPH radical scavenging and ferric ion-reducing activities in a dose-dependent manner. Cyanidin-3-rutinoside, the most abundant polyphenol compound of MAJ identified by LC-MS in this study, showed weak inhibitory effects against FL04 in the pretreatment, whereas gallic acid, a minor compound of MAJ, revealed significant antiviral effect. These results suggest that MAJ can be developed as a novel plant-derived antiviral against influenza viruses.

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