Murine C-type RNA Virus from Spontaneous Neoplasms: in vitro Host Range and Oncogenic Potential

MEX3A belongs to the MEX3 (Muscle EXcess) protein family consisting of four members (MEX3A-D) in humans. Characteristic for MEX3 proteins is their domain structure with 2 HNRNPK homology (KH) domains mediating RNA binding and a C-terminal really interesting new gene (RING) domain that harbors E3 ligase function. In agreement with their domain composition, MEX3 proteins were reported to modulate both RNA fate and protein ubiquitination. MEX3 paralogs exhibit an oncofetal expression pattern, they are severely downregulated postnatally, and re-expression is observed in various malignancies. Enforced expression of MEX3 proteins in various cancers correlates with poor prognosis, emphasizing their oncogenic potential. The latter is supported by MEX3A’s impact on proliferation, self-renewal as well as migration of tumor cells in vitro and tumor growth in xenograft studies.

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Revealing biophysical properties of KfrA-type proteins as a novel class of cytoskeletal, coiled-coil plasmid-encoded proteins

BMC Microbiology ◽

10.1186/s12866-020-02079-w ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

M. Adamczyk ◽

E. Lewicka ◽

R. Szatkowska ◽

H. Nieznanska ◽

J. Ludwiczak ◽

...

Keyword(s):

Dna Binding ◽

Host Range ◽

Coiled Coil ◽

Amino Acid Sequences ◽

Broad Host Range ◽

Biophysical Properties ◽

Dna Binding Sites ◽

In Silico Studies ◽

Wide Range

Abstract Background DNA binding KfrA-type proteins of broad-host-range bacterial plasmids belonging to IncP-1 and IncU incompatibility groups are characterized by globular N-terminal head domains and long alpha-helical coiled-coil tails. They have been shown to act as transcriptional auto-regulators. Results This study was focused on two members of the growing family of KfrA-type proteins encoded by the broad-host-range plasmids, R751 of IncP-1β and RA3 of IncU groups. Comparative in vitro and in silico studies on KfrAR751 and KfrARA3 confirmed their similar biophysical properties despite low conservation of the amino acid sequences. They form a wide range of oligomeric forms in vitro and, in the presence of their cognate DNA binding sites, they polymerize into the higher order filaments visualized as “threads” by negative staining electron microscopy. The studies revealed also temperature-dependent changes in the coiled-coil segment of KfrA proteins that is involved in the stabilization of dimers required for DNA interactions. Conclusion KfrAR751 and KfrARA3 are structural homologues. We postulate that KfrA type proteins have moonlighting activity. They not only act as transcriptional auto-regulators but form cytoskeletal structures, which might facilitate plasmid DNA delivery and positioning in the cells before cell division, involving thermal energy.

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Investigation of Salmonella Phage–Bacteria Infection Profiles: Network Structure Reveals a Gradient of Target-Range from Generalist to Specialist Phage Clones in Nested Subsets

Viruses ◽

10.3390/v13071261 ◽

2021 ◽

Vol 13 (7) ◽

pp. 1261

Author(s):

Khatuna Makalatia ◽

Elene Kakabadze ◽

Nata Bakuradze ◽

Nino Grdzelishvili ◽

Ben Stamp ◽

...

Keyword(s):

Host Range ◽

Salmonella Enterica ◽

Target Range ◽

Nested Subsets ◽

Specific Phage ◽

Salmonella Infections ◽

Specialist Species ◽

Host Infection ◽

Salmonella Phage

Bacteriophages that lyse Salmonella enterica are potential tools to target and control Salmonella infections. Investigating the host range of Salmonella phages is a key to understand their impact on bacterial ecology, coevolution and inform their use in intervention strategies. Virus–host infection networks have been used to characterize the “predator–prey” interactions between phages and bacteria and provide insights into host range and specificity. Here, we characterize the target-range and infection profiles of 13 Salmonella phage clones against a diverse set of 141 Salmonella strains. The environmental source and taxonomy contributed to the observed infection profiles, and genetically proximal phages shared similar infection profiles. Using in vitro infection data, we analyzed the structure of the Salmonella phage–bacteria infection network. The network has a non-random nested organization and weak modularity suggesting a gradient of target-range from generalist to specialist species with nested subsets, which are also observed within and across the different phage infection profile groups. Our results have implications for our understanding of the coevolutionary mechanisms shaping the ecological interactions between Salmonella phages and their bacterial hosts and can inform strategies for targeting Salmonella enterica with specific phage preparations.

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Exploration of the in vitro cytotoxic and antiviral activities of different medicinal plants against infectious bursal disease (IBD) virus

Open Life Sciences ◽

10.2478/s11535-013-0276-8 ◽

2014 ◽

Vol 9 (5) ◽

pp. 531-542 ◽

Cited By ~ 1

Author(s):

Waqas Ahmad ◽

Sohail Ejaz ◽

Khaleeq Anwar ◽

Muhammad Ashraf

Keyword(s):

Medicinal Plants ◽

Colorimetric Assay ◽

Rna Virus ◽

Infectious Bursal Disease ◽

Dna Viruses ◽

Mtt Colorimetric Assay ◽

Bursal Disease ◽

Dried Fruit ◽

Antiviral Activities

AbstractInfectious bursal disease (IBD) caused by non-enveloped double stranded RNA virus is an acute and contagious poultry disease. Outbreak of IBD could result in 10–75% mortality of the birds; hence it has gained socio-economic importance worldwide. Medicinal plants have shown broad spectrum anti-viral activities against RNA and DNA viruses. Moringa oleifera Lam (MOL), Phyllanthus emblicus Linn (PEL), Glycyrrhiza glabra Linn (GGL), and Eugenia jambolana Lam (EJL) are commonly available medicinal plants of the sub-continent and exhibited anti-viral potential against different viruses. Ethanolic extracts of the leaves of MOL and EJL, roots of GGL and dried fruit of PEL were investigated for their cytotoxic and anti-viral potential against IBD virus using MTT colorimetric assay and anti-viral assay. Significant anti-viral potential (P<0.001) was demonstrated at concentrations 12.5, 25, 50 and 100 µg ml−1 of GGL, PEL, MOL and EJL, respectively, with no cytotoxicity. Data also spotlighted that all tested plant extracts possess significant anti-viral potential and this trend was higher in GGL followed by PEL, MOL, and EJL. The data undoubtedly conclude that these medicinal plants contain several health beneficial phyto-chemicals which got significant anti-viral potential and effectively be utilized against IBD virus. Moreover, the outcomes of this study provide a platform on the way to discover novel anti-viral agents against IBD virus and other viruses from plant origin.

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Biocontrol of seed-borne Alternaria raphani and A. brassicicola

Canadian Journal of Microbiology ◽

10.1139/m87-149 ◽

1987 ◽

Vol 33 (10) ◽

pp. 850-856 ◽

Cited By ~ 27

Author(s):

G. Vannacci ◽

G. E. Harman

Keyword(s):

Biological Control ◽

Host Range ◽

Trichoderma Harzianum ◽

Pythium Species ◽

Biological Control Agents ◽

Fusarium Sp ◽

Chamber Studies ◽

Growth Chamber Studies ◽

Radish Seedlings

Forty-two microorganisms were tested as biological control agents against Alternaria raphani and A. brassicicola. Tests were conducted for in vitro antagonistic ability, for ability to control the pathogens on naturally infected seeds germinated on moistened blotters, and in planting mix in growth chamber studies, and for their ability to reduce pod infection. The organisms tested were obtained from cruciferous seeds or were strains already identified as being effective against soil-borne Pythium species. The blotter test indicated that six organisms increased both the number of healthy seedlings and the number of seedlings produced from A. raphani infected radish seeds. An additional seven strains improved either germination or increased the number of healthy seedlings. Twenty-nine organisms increased the number of healthy cabbage seedlings from A. brassicicola infected seeds, but total germination was not modified by any treatment. Experiments in planting mix showed that five antagonists (Chaetomium globosum, two strains of Trichoderma harzianum, T. koningii, and Fusarium sp.) increased the number of healthy plants in both radish samples tested, while four additional antagonists provided a significant increase in only one of the samples tested. The five antagonists that consistently increased numbers of healthy radish seedlings also decreased pod infection by A. raphani. None were as effective as iprodrone, however. Several effective antagonists were found to be mycoparasitic against Alternaria spp. Some strains of Trichoderma previously found to be effective against Pythium spp. were also effective against Alternaria spp., indicating that these strains have a wide host range.

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Glycoprotein D-Independent Infectivity of Pseudorabies Virus Results in an Alteration of In Vivo Host Range and Correlates with Mutations in Glycoproteins B and H

Journal of Virology ◽

10.1128/jvi.75.21.10054-10064.2001 ◽

2001 ◽

Vol 75 (21) ◽

pp. 10054-10064 ◽

Cited By ~ 18

Author(s):

Jerg Schmidt ◽

Volker Gerdts ◽

Jörg Beyer ◽

Barbara G. Klupp ◽

Thomas C. Mettenleiter

Keyword(s):

Host Range ◽

Pseudorabies Virus ◽

Natural Host ◽

Target Cells ◽

Glycoprotein D ◽

Wild Type ◽

Cellular Receptors ◽

Receptor Usage

ABSTRACT Infection of cells by herpesviruses is initiated by the interaction of viral envelope glycoproteins with cellular receptors. In the alphaherpesvirus pseudorabies virus (PrV), the causative agent of Aujeszky's disease in pigs, the essential glycoprotein D (gD) mediates secondary attachment of virions to target cells by binding to newly identified cellular receptors (R. J. Geraghty, C. Krummenacher, G. H. Cohen, R. J. Eisenberg, and P. G. Spear, Science 280:1618–1620, 1998). However, in the presence of compensatory mutations, infection can also occur in the absence of gD, as evidenced by the isolation in cell culture of an infectious gD-negative PrV mutant (PrV-gD− Pass) (J. Schmidt, B. G. Klupp, A. Karger, and T. C. Mettenleiter, J. Virol. 71:17–24, 1997). PrV-gD− Pass is replication competent with an only moderate reduction in specific infectivity but appears to bind to receptors different from those recognized by wild-type PrV (A. Karger, J. Schmidt, and T. C. Mettenleiter, J. Virol. 72:7341–7348, 1998). To analyze whether this alteration in receptor usage in vitro influences infection in vivo, the model host mouse and the natural host pig were intranasally infected with PrV-gD− Pass and were compared to animals infected by wild-type PrV. For mice, a comparable progress of disease was observed, and all animals infected with mutant virus died, although they exhibited a slight delay in the onset of symptoms and, correspondingly, a longer time to death. In contrast, whereas wild-type PrV-infected pigs showed clinical signs and histological and histopathological findings typical of PrV infection, no signs of disease were observed after infection with PrV-gD− Pass. Moreover, in these animals, virus-infected cells were not detectable by immunohistochemical staining of different organ samples and no virus could be isolated from nasal swabs. Mutations in glycoproteins B and H were found to correlate with, and probably contribute to, gD-independent infectivity. In conclusion, although PrV-gD− Pass is virulent in mice, it is apparently unable to infect the natural host, the pig. This altered host range in vivo correlates with a difference of receptor usage in vitro and demonstrates for the first time the importance of gD receptors in alphaherpesvirus infection of an animal host.

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Liposomal Systems as Nanocarriers for the Antiviral Agent Ivermectin

International Journal of Biomaterials ◽

10.1155/2016/8043983 ◽

2016 ◽

Vol 2016 ◽

pp. 1-15 ◽

Cited By ~ 11

Author(s):

Romina Croci ◽

Elisabetta Bottaro ◽

Kitti Wing Ki Chan ◽

Satoru Watanabe ◽

Margherita Pezzullo ◽

...

Keyword(s):

Yellow Fever ◽

Rna Virus ◽

Multiorgan Failure ◽

Antiviral Agent ◽

Drug Carriers ◽

Virus Infections ◽

Starting Point ◽

Significant Public Health ◽

High Cytotoxicity

RNA virus infections can lead to the onset of severe diseases such as fever with haemorrhage, multiorgan failure, and mortality. The emergence and reemergence of RNA viruses continue to pose a significant public health threat worldwide with particular attention to the increasing incidence of flaviviruses, among others Dengue, West Nile Virus, and Yellow Fever viruses. Development of new and potent antivirals is thus urgently needed. Ivermectin, an already known antihelminthic drug, has shown potent effectsin vitroonFlavivirushelicase, with EC50values in the subnanomolar range for Yellow Fever and submicromolar EC50for Dengue Fever, Japanese encephalitis, and tick-borne encephalitis viruses. However ivermectin is hampered in its application by pharmacokinetic problems (little solubility and high cytotoxicity). To overcome such problems we engineered different compositions of liposomes as ivermectin carriers characterizing and testing them on several cell lines for cytotoxicity. The engineered liposomes were less cytotoxic than ivermectin alone and they showed a significant increase of the antiviral activity in all the Dengue stains tested (1, 2, and S221). In the current study ivermectin is confirmed to be an effective potential antiviral and liposomes, as drug carriers, are shown to modulate the drug activity. All together the results represent a promising starting point for future improvement of ivermectin as antiviral and its delivery.

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Advancing RNA Virus Discovery and Biology with Whole Genome Sequencing

10.21007/etd.cghs.2021.0551 ◽

2021 ◽

Author(s):

◽

Mariah Taylor ◽

Keyword(s):

Genetic Variation ◽

Amino Acid ◽

Whole Genome Sequencing ◽

Genome Sequencing ◽

Rna Virus ◽

Animal Health ◽

Functional Domains ◽

Whole Genome ◽

Coding Region

Two RNA virus families that pose a threat to human and animal health are Hantaviridae and Coronaviridae. These RNA viruses which originate in wildlife continue and will continue to cause disease, and hence, it is critical that scientific research define the mechanisms as to how these viruses spillover and adapt to new hosts to become endemic. One gap in our ability to define these mechanisms is the lack of whole genome sequences for many of these viruses. To address this specific gap, I developed a versatile amplicon-based whole-genome sequencing (WGS) approach to identify viral genomes of hantaviruses and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) within reservoir and spillover hosts. In my research studies, I used the amplicon-based WGS approach to define the genetic plasticity of viral RNA within pathogenic and nonpathogenic hantavirus species. The standing genetic variation of Andes orthohantavirus and Prospect Hill orthohantavirus was mapped out and amino acid changes occurring outside of functional domains were identified within the nucleocapsid and glycoprotein. I observed several amino acid changes in functional domains of the RNA-dependent RNA polymerase, as well as single nucleotide polymorphisms (SNPs) within the 3’ non-coding region (NCR) of the S-segment. To identify whether virus adaptation would occur within the S- and L-segments we attempted to adapt hantaviruses in vitro in a spillover host model through passaging experiments. In early passages we identified few mutations in the M-segment with the majority being identified in the S-segment 3’ NCR and the L-segment. This work suggests that hantavirus adaptation occurs in the S- and L-segments although the effect of these mutants on pathology is yet to be determined. While sequencing laboratory isolates is easily accomplished, sequencing low concentrations of virus within the reservoir is a formidable task. I further translated our amplicon-based WGS approach into a pan-oligonucleotide amplicon-based WGS approach to sequence hantavirus vRNA and mRNA from reservoir and spillover hosts in Ukraine. This approach successfully identified a novel Puumala orthohantavirus (PUUV) strain in Ukraine and using Bayesian phylogenetics we found this strain to be associated with the PUUV Latvian lineage. Early during the SARS-CoV-2 pandemic, I applied the knowledge gained in the hantavirus WGS efforts to sequencing of SARS-CoV-2 from nasopharyngeal swabs collected in April 2020. The genetic diversity of 45 SARS-CoV-2 isolates was evaluated with the methods I developed. We identified D614G, a notable mutation known for increasing transmission, in over 90% of our isolates. Two major lineages distinguish SARS-CoV-2 variants worldwide, lineages A and B. While most of our isolates were found within B lineage, we also identified one isolate within lineage A. We performed in vitro work which confirmed A lineage isolates as having poor replication in the trachea as compared to the nasal cavity. Five of these isolates presented a unique array of mutations which were assessed in the keratin 18 human angiotensin-converting enzyme 2 (K18-hACE2) mouse model for its response immunologically and pathogenically. We identified a distinction of pathogenesis between the A and B lineages with emphysema being common amongst A lineage isolates. Additionally, we discovered a small cohort of likely SNPs that defined the late induction of eosinophils during infection. In summary, this work will further define the dynamics of genetic variation and plasticity within virus populations that cause disease outbreaks and will allow a deeper understanding of the virus-host relationship.

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TaffiX® nasal powder spray forms an effective barrier against infectious new variants of SARS-CoV-2 (COVID-19)

10.21203/rs.3.rs-420365/v1 ◽

2021 ◽

Author(s):

Michal Mandelboim ◽

Ella Mendelson ◽

Yaron Drori ◽

Nofar Atari ◽

Tair Lapidot ◽

...

Keyword(s):

South African ◽

Rna Virus ◽

Rna Extraction ◽

Real Life ◽

Preclinical Study ◽

Viral Rna ◽

Pcr Analysis ◽

Effective Barrier ◽

Gel Layer

Abstract Introduction: While vaccination efforts against SARS-CoV-2 around the world are ongoing -, new high-infectious variants of the virus are being detected. The protection of the available vaccines against some of the new variants is weaker, and experts are concerned that newer as yet undescribed variants of this mutated RNA virus will eventually prove stable against the current vaccines. Additional preventive measures will therefore be needed to protect the population until effective vaccinations are widely available.TaffiX® is a personal, anti-viral nasal powder spray comprised of low pH Hypromellose that upon insufflation into the nose creates a thin gel layer covering the nasal mucosa and forming a protective mechanical barrier that prevents viruses from engaging with nasal cells- the main portal of entry for viruses. Taffix is commercially available in many countries across Europe, Asia America and Africa. In a prior preclinical study, TaffiX® was found to be effective against SARS-CoV-2 Hong Kong/VM20001061/2020 in experimental in vitro conditions. A real-life clinical survey demonstrated that TaffiX® nasal spray significantly reduced the SARS-CoV-2 infection rate post mass-gathering event in a highly endemic community.Objective: The current study aimed to test the protective effect of Taffix against new pathogenic, highly infectious SARS-CoV-2 variants in vitro: the “British” B.1.1.7 (hCoV-19/Israel/CVL-46879-ngs/2020) and the “South African” B.1.351 (hCoV-19/Israel/CVL-2557-ngs/2020) variants.Study design: A TaffiX® gel was formed on a nylon filter, using an amount equivalent to a clinical dose of Taffix . Filters were then seeded with SARS-CoV-2 B.1.1.7 (“British”) and B.1.351 (“South African”) variants. After a 10 -minute incubation at room temperature, the bottom of each filter was washed, and the resulting flow-through was collected and seeded into 24 -well plates containing Vero-E6 cells. After 5 days of incubation, a 200 µl sample from each well was taken for viral RNA extraction followed by SARS-CoV 2 RT-PCR analysis.Results: The TaffiX® gel completely blocked SARS-CoV-2 highly infectious variants B.1.1.7 and B.1.351 in vitro, reducing the titer of recoverable infectious virus as well as viral RNA by 100%.Conclusions: Under in vitro conditions, TaffiX® formed an effective protective barrier against SARS-COV-2 variants (British variant and South African Variant). These results are consistent with prior findings demonstrating the in vitro high efficacy of Taffix gel in preventing viruses from reaching cells and infecting them. These results, added to clinical real-life studies performed with Taffix , support its use as an effective barrier against new variants of SARS-CoV-2 in conjunction with other protective measures.

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