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      6-methylmercaptopurine riboside, a thiopurine nucleoside with antiviral activity against canine distemper virus in vitro

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          Abstract

          Background

          Canine distemper (CD) is a widespread infectious disease that can severely impact a variety of species in the order Carnivora, as well as non-carnivore species such as non-human primates. Despite large-scale vaccination campaigns, several fatal outbreaks have been reported in wild and domestic carnivore populations. This, in association with expansion of the disease host range and the development of vaccine-escape strains, has contributed to an increased demand for therapeutic strategies synergizing with vaccine programs for effectively controlling canine distemper. 6-methylmercaptopurine riboside (6MMPr) is a modified thiopurine nucleoside with known antiviral properties against certain RNA viruses.

          Methods

          We tested the inhibitory effects of 6MMPr against a wild-type CDV strain infection in cell culture. We measured infectious particle production and viral RNA levels in treated and untreated CDV-infected cells. Ribavirin (RIB) was used as a positive control.

          Results

          Here, we report for the first time the antiviral effects of 6MMPr against canine distemper virus (CDV) in vitro. 6MMPr was able to reduce viral RNA levels and to inhibit the production of infectious CDV particles. The therapeutic selectivity of 6MMPr was approximately six times higher than that of ribavirin.

          Conclusion

          Our results indicate that 6MMPr has high anti-CDV potential and warrants further testing against other paramyxoviruses, as well as clinical testing of the compound against CDV.

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          Most cited references20

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          Cross-species transmission of canine distemper virus—an update

          Canine distemper virus (CDV) is a pantropic morbillivirus with a worldwide distribution, which causes fatal disease in dogs. Affected animals develop dyspnea, diarrhea, neurological signs and profound immunosuppression. Systemic CDV infection, resembling distemper in domestic dogs, can be found also in wild canids (e.g. wolves, foxes), procyonids (e.g. raccoons, kinkajous), ailurids (e.g. red pandas), ursids (e.g. black bears, giant pandas), mustelids (e.g. ferrets, minks), viverrids (e.g. civets, genets), hyaenids (e.g. spotted hyenas), and large felids (e.g. lions, tigers). Furthermore, besides infection with the closely related phocine distemper virus, seals can become infected by CDV. In some CDV outbreaks including the mass mortalities among Baikal and Caspian seals and large felids in the Serengeti Park, terrestrial carnivores including dogs and wolves have been suspected as vectors for the infectious agent. In addition, lethal infections have been described in non-carnivore species such as peccaries and non-human primates demonstrating the remarkable ability of the pathogen to cross species barriers. Mutations affecting the CDV H protein required for virus attachment to host-cell receptors are associated with virulence and disease emergence in novel host species. The broad and expanding host range of CDV and its maintenance within wildlife reservoir hosts considerably hampers disease eradication.
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            Detection of canine distemper virus nucleoprotein RNA by reverse transcription-PCR using serum, whole blood, and cerebrospinal fluid from dogs with distemper.

            Reverse transcription-PCR (RT-PCR) was used to detect canine distemper virus (CDV) nucleoprotein (NP) RNA in serum, whole blood, and cerebrospinal fluid (CSF) samples from 38 dogs with clinically suspected distemper. Results were correlated to clinical findings, anti-CDV neutralizing antibody titers, postmortem findings, and demonstration of CDV NP antigen by immunohistochemistry. The specificity of the RT-PCR was ensured by amplification of RNA from various laboratory CDV strains, restriction enzyme digestion, and Southern blot hybridization. In 29 of 38 dogs, CDV infection was confirmed by postmortem examination and immunohistochemistry. The animals displayed the catarrhal, systemic, and nervous forms of distemper. Seventeen samples (serum, whole blood, or CSF) from dogs with distemper were tested with three sets of primers targeted to different regions of the NP gene of the CDV Onderstepoort strain. Expected amplicons were observed in 82, 53, and 41% of the 17 samples, depending upon the primer pair used. With the most sensitive primer pair (primer pair I), CDV NP RNA was detected in 25 of 29 (86%) serum samples and 14 of 16 (88%) whole blood and CSF samples from dogs with distemper but not in body fluids from immunohistochemically negative dogs. Nucleotide sequence analysis of five RT-PCR amplicons from isolates from the field revealed few silent point mutations. These isolates exhibited greater homology to the Rockborn (97 to 99%) than to the Onderstepoort (95 to 96%) CDV strain. In summary, although the sensitivity of the RT-PCR for detection of CDV is strongly influenced by the location of the selected primers, this nucleic acid detection system represents a highly specific and sensitive method for the antemortem diagnosis of distemper in dogs, regardless of the form of distemper, humoral immune response, and viral antigen distribution.
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              The RNA synthesis machinery of negative-stranded RNA viruses.

              The group of Negative-Stranded RNA Viruses (NSVs) includes many human pathogens, like the influenza, measles, mumps, respiratory syncytial or Ebola viruses, which produce frequent epidemics of disease and occasional, high mortality outbreaks by transmission from animal reservoirs. The genome of NSVs consists of one to several single-stranded, negative-polarity RNA molecules that are always assembled into mega Dalton-sized complexes by association to many nucleoprotein monomers. These RNA-protein complexes or ribonucleoproteins function as templates for transcription and replication by action of the viral RNA polymerase and accessory proteins. Here we review our knowledge on these large RNA-synthesis machines, including the structure of their components, the interactions among them and their enzymatic activities, and we discuss models showing how they perform the virus transcription and replication programmes.
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                Author and article information

                Contributors
                otaviovet@gmail.com
                mf-dany@hotmail.com
                cctozato@gmail.com
                jufietto@ufv.br
                marcia@ufv.br
                gustavo.bressan@ufv.br
                55 81 2123-7849 , lindomar.pena@cpqam.fiocruz.br
                55 31 3899-1471 , abelardo.junior@ufv.br
                Journal
                Virol J
                Virol. J
                Virology Journal
                BioMed Central (London )
                1743-422X
                26 June 2017
                26 June 2017
                2017
                : 14
                : 124
                Affiliations
                [1 ]ISNI 0000 0000 8338 6359, GRID grid.12799.34, Laboratory of Animal Virology, Department of Veterinary, , Universidade Federal of Viçosa, ; Av. Peter Henry Rolfs, s/n, Viçosa, MG 36570-000 Brazil
                [2 ]ISNI 0000 0001 0723 0931, GRID grid.418068.3, , Laboratory of Virology and Experimental Therapy, Oswaldo Cruz Foundation (FIOCRUZ), Aggeu Magalhães Research Center, ; Av. Moraes Rego, s/n, Campus UFPE, Cidade Universitária, Recife, PE 50670-420 Brazil
                [3 ]ISNI 0000 0001 2188 478X, GRID grid.410543.7, Laboratory of Animal and Human Virology, Department of Microbiology and Immunology, Biosciences Institute, , Paulista State University, ; Botucatu, SP 18618-970 Brazil
                [4 ]ISNI 0000 0000 8338 6359, GRID grid.12799.34, Department of Biochemistry and Molecular Biology, , Federal University of Viçosa, ; Av. Peter Henry Rolfs, s/n, Viçosa, MG 36570-000 Brazil
                Author information
                http://orcid.org/0000-0002-0614-7527
                Article
                785
                10.1186/s12985-017-0785-6
                5485505
                28651549
                49da4754-b539-4ae5-8129-65e1c6b92d8c
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 3 October 2016
                : 12 June 2017
                Funding
                Funded by: Coordination for the Improvement of Higher Education Personnel - CAPES
                Award ID: 23038.004678/2015-24
                Award Recipient :
                Funded by: National Council for Scientific and Technological Development - CNPq
                Award ID: 304727/2016-4
                Funded by: Minas Gerais State Research Foundation - FAPEMIG
                Award ID: PPM-00796-15
                Categories
                Research
                Custom metadata
                © The Author(s) 2017

                Microbiology & Virology
                canine distemper,antiviral,azathioprine,thiopurine,nucleoside analogue
                Microbiology & Virology
                canine distemper, antiviral, azathioprine, thiopurine, nucleoside analogue

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