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Brazilian Journal of Microbiology :... Jan 2010Bovine herpesvirus type 5 (BoHV-5) is the agent of meningoencephalitis, an important disease of cattle in South America. The neuropathogenesis of BoHV-5 infection is...
Bovine herpesvirus type 5 (BoHV-5) is the agent of meningoencephalitis, an important disease of cattle in South America. The neuropathogenesis of BoHV-5 infection is poorly understood and most previous research focused on the role of envelope glicoproteins in neurovirulence. Thymidine kinase (TK) is a viral enzyme necessary for virus replication in neurons and, therefore, represents a potential target for virus attenuation. The selection and characterization of BoHV-5 variants resistant to the nucleoside analog brivudin (BVDU), which selects TK-defective viruses is here described. Several BVDU-resistant clones were obtained after multiple passages in tissue culture in the presence of BVDU and one clone (BoHV-5/R-27) was further characterized. The selected clone replicated to similar titers and produced plaques with similar size and morphology to those of wild-type virus (SV507/99). The genetic stability of the resistant virus was demonstrated after ten passages in cell culture in the absence of the drug. Moreover, the drug-resistant virus showed reduced virulence in a rabbit model: virus inoculation in four rabbits did not result in disease, in contrast with 75% morbidity (3/4) and 50% mortality (2/2) among rabbits inoculated with the parental virus. These results demonstrate that BoHV-5 is sensitive to BVDU and that drug-resistant mutants can be readily selected upon BVDU treatment. BVDU-resistant mutants, likely defective in TK, retained their ability to replicate in tissue culture yet were attenuated for rabbits. This strategy to obtain TK-defective BoHV-5 may be useful to study the role of TK in BoHV-5 neuropathogenesis and for vaccine development.
PubMed: 24031472
DOI: 10.1590/S1517-838220100001000019 -
Acta Dermato-venereologica Mar 2014Herpes zoster neural injury was assessed by determining cutaneous nerve density in skin biopsies from the affected dermatomes of 35 adult patients with herpes zoster in... (Randomized Controlled Trial)
Randomized Controlled Trial
Herpes zoster neural injury was assessed by determining cutaneous nerve density in skin biopsies from the affected dermatomes of 35 adult patients with herpes zoster in the acute phase and 3 months post-treatment, using protein gene product 9.5 immunohistochemistry. In contrast to the significant increase in subepidermal nerve fibre density (11.77 ± 4.88/mm vs. 13.29 ± 5.74/mm, p = 0.045) after 3 months, no differences were found in epidermal free nerve endings (2.43 ± 2.35/mm and 2.8 ± 2.86/mm, p = 0.168). Patients with post-herpetic neuralgia had significantly lower subepidermal nerve fibre densities (9.7 ± 2.05/mm vs. 14.72 ± 6.13/mm, p = 0.011) compared with non-post-herpetic neuralgia patients. No differences in cutaneous nerve density were found in relation to antiviral therapy. In conclusion, 3 months after acute infection, no sign of epidermal innervation recovery is observed, while the increased subepidermal nerve fibre density in the affected dermatomes probably reflects nerve regeneration that is not affected by antiviral agent type. Subepidermal nerve fibre density is decreased in patients with post-herpetic neuralgia 3-months post-acute herpes zoster infection.
Topics: 2-Aminopurine; Acyclovir; Antiviral Agents; Biomarkers; Biopsy; Bromodeoxyuridine; Famciclovir; Female; Herpes Zoster; Humans; Immunohistochemistry; Male; Middle Aged; Nerve Fibers; Neuralgia, Postherpetic; Skin; Ubiquitin Thiolesterase; Valacyclovir; Valine
PubMed: 23995395
DOI: 10.2340/00015555-1664 -
Antiviral Chemistry & Chemotherapy 2006Previous studies have shown that cycloSaligenyl-monophosphate (cycloSal-MP) derivatives of aciclovir (ACV), penciclovir (PCV) and brivudin (BVDU) can act as inhibitors...
Previous studies have shown that cycloSaligenyl-monophosphate (cycloSal-MP) derivatives of aciclovir (ACV), penciclovir (PCV) and brivudin (BVDU) can act as inhibitors of vaccinia virus and cowpox virus replication in vitro. The aim of the present study was to evaluate the inhibatory efficacy on DNA synthesis in vaccinia and cowpox viruses of several cycloSal-pro-nucleotides of ACV and BVDU, which have proven activity against pox viruses. Viral DNA was quantified in treated and non-treated virus-infected cells by semi-quantitative PCR on the basis of the haemagglutinin protein gene of orthopoxviruses. As result, an inhibitory efficacy on vaccinia and cowpox virus DNA replication could be demonstrated for 3-methyl-cycloSal-ACVMP, 5-H-cycloSal-ACVMP, 6-chloro-7-ECM-cycloSal-3'-OH-BVDUMP, and 6-chloro-7-methyl-cycloSal-3'-OH-BVDUMP. At concentrations of 32-128 mg/ml, 3-methyl-cyc/oSal-ACVMP and 6-chloro-7-ECM-cycloSal-3'OH-BVDUMP inhibited synthesis of viral DNA to a similar extent as the well-known inhibitors of pox viruses, cidofovir and 5-iodo-dUrd (deoxyuridine). When concentrations of 128 mg/ml were administered, both test substances diminished the amount of viral genome copies by > or =4 log10 corresponding to > or =99.99% reduction. In conclusion, selected cycloSal-pro-nucleotide derivatives of ACV and BVDU can inhibit orthopoxviral DNA synthesis. The high inhibitory efficacy on both replication of viral DNA and infectious viral particles in cell cultures makes these compounds promising candidates for in vivo experiments.
Topics: Acyclovir; Animals; Antiviral Agents; Bromodeoxyuridine; Chlorocebus aethiops; DNA Replication; DNA, Viral; Dose-Response Relationship, Drug; Orthopoxvirus; Polymerase Chain Reaction; Poxviridae Infections; Vero Cells; Virus Replication
PubMed: 16542003
DOI: 10.1177/095632020601700104 -
The FEBS Journal May 2008The Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (dNK; EC 2.7.1.145) has a high turnover rate and a wide substrate range that makes it a very good...
The Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (dNK; EC 2.7.1.145) has a high turnover rate and a wide substrate range that makes it a very good candidate for gene therapy. This concept is based on introducing a suicide gene into malignant cells in order to activate a prodrug that eventually may kill the cell. To be able to optimize the function of dNK, it is vital to have structural information of dNK complexes. In this study we present crystal structures of dNK complexed with four different nucleoside analogs (floxuridine, brivudine, zidovudine and zalcitabine) and relate them to the binding of substrate and feedback inhibitors. dCTP and dGTP bind with the base in the substrate site, similarly to the binding of the feedback inhibitor dTTP. All nucleoside analogs investigated bound in a manner similar to that of the pyrimidine substrates, with many interactions in common. In contrast, the base of dGTP adopted a syn-conformation to adapt to the available space of the active site.
Topics: Adenosine Diphosphate; Animals; Antimetabolites; Bromodeoxyuridine; Cytarabine; Cytidine Triphosphate; Drosophila Proteins; Drosophila melanogaster; Feedback; Floxuridine; Guanosine Triphosphate; Hydrogen Bonding; Inhibitory Concentration 50; Kinetics; Models, Chemical; Models, Molecular; Phosphotransferases (Alcohol Group Acceptor); Protein Binding; Protein Structure, Secondary; Structure-Activity Relationship; Thymine Nucleotides; X-Ray Diffraction; Zalcitabine; Zidovudine
PubMed: 18384378
DOI: 10.1111/j.1742-4658.2008.06369.x -
Microbiology and Immunology 1988The highly potent and selective anti-herpesvirus agent, (E)-5-(2-bromovinyl)-2'deoxyuridine (BVdU), was examined for its inhibitory effect on the salmonid herpesviruses...
The highly potent and selective anti-herpesvirus agent, (E)-5-(2-bromovinyl)-2'deoxyuridine (BVdU), was examined for its inhibitory effect on the salmonid herpesviruses Oncorhynchus masou virus (OMV) and Herpesvirus salmonis (H. salmonis). Minimum inhibitory concentrations (MIC) of BVdU for OMV and H. salmonis were 1.25 and 3.0 micrograms/ml, respectively; these values were equal to or higher than those obtained for acyclovir or cytarabine. OMV DNA polymerase activity was reduced in a dose-dependent fashion by BVdU 5'-triphosphate (BVdUTP) within the concentration range of 3 to 30 microM. However, BVdUTP could also be substituted for the natural substrate, TTP, in the OMV DNA polymerase assay. It is postulated that the inhibitory action of BVdU on the salmonid herpesviruses is more or less similar to that on other herpesviruses and resides with respect to the inhibition of the virus DNA polymerase activity as well as incorporation of BVdU into the viral DNA.
Topics: Acyclovir; Animals; Antiviral Agents; Bromodeoxyuridine; Cells, Cultured; DNA-Directed DNA Polymerase; Gonads; Herpesviridae; Salmonidae; Vidarabine; Virus Replication
PubMed: 3374404
DOI: 10.1111/j.1348-0421.1988.tb01365.x -
Journal of Virology May 2005A broad variety of herpes simplex virus type 1 clones was selected under a single round of high-dose selection with brivudin. Mutations in the thymidine kinase (TK)...
A broad variety of herpes simplex virus type 1 clones was selected under a single round of high-dose selection with brivudin. Mutations in the thymidine kinase (TK) genes consisted of 42% frameshift mutations within homopolymer repeats of G's and C's and single nucleotide substitutions (58%) that produced stop codons (Q261 and R281) or a new codon at the site of the substitution (A168T, R51W, G59W, G206R, R220H, Y239S, and T287 M). The A168T change, associated with an altered TK phenotype, proved to be the most commonly selected substitution. For the different mutants, a correlation between phenotype, genotype, and in vivo neurovirulence was observed.
Topics: Animals; Antiviral Agents; Bromodeoxyuridine; Chlorocebus aethiops; Codon, Terminator; Drug Resistance, Viral; Frameshift Mutation; Models, Molecular; Phenotype; Selection, Genetic; Simplexvirus; Thymidine Kinase; Vero Cells
PubMed: 15827202
DOI: 10.1128/JVI.79.9.5863-5869.2005 -
Japanese Journal of Cancer Research :... Apr 1990The antitumor activity of 5'-deoxy-5-fluorouridine (DFUR), a prodrug of 5-fluorouracil (5-FU), is markedly enhanced if DFUR treatment is combined with...
The antitumor activity of 5'-deoxy-5-fluorouridine (DFUR), a prodrug of 5-fluorouracil (5-FU), is markedly enhanced if DFUR treatment is combined with (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU). Combined oral administration of DFUR (10 mg/kg) and BVDU (10 mg/kg) three times (every 3 h) per day for 5 days afforded greater antitumor activity than a single dose of DFUR (300 mg/kg/day) for 5 days in mice bearing either adenocarcinoma 755 or Lewis lung carcinoma, while in the colon 26 system the antitumor effects of both treatment regimens were equivalent. Thus, a low-dose regimen of DFUR when combined with BVDU provides a similar or greater antitumor activity than a high-dose regimen of DFUR that is not combined with BVDU. The area under the curve of plasma 5-FU following a treatment with the combination of DFUR (10 mg/kg) and BVDU (10 mg/kg) was equal to that following DFUR (300 mg/kg) treatment.
Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Antiviral Agents; Bromodeoxyuridine; Colonic Neoplasms; Drug Therapy, Combination; Floxuridine; Fluorouracil; Liver Neoplasms; Lung Neoplasms; Mice; Mice, Inbred BALB C; Pentosyltransferases; Prodrugs; Pyrimidine Phosphorylases
PubMed: 2142152
DOI: 10.1111/j.1349-7006.1990.tb02586.x -
Microbiology and Immunology 1996We developed a rapid and simple method for the screening of antiviral agents against herpes simplex virus (HSV) in a model of gastrointestinal herpetic infection in...
We developed a rapid and simple method for the screening of antiviral agents against herpes simplex virus (HSV) in a model of gastrointestinal herpetic infection in vitro. This method was based on inhibition of HSV-induced cytopathogenicity in gastric adenocarcinoma MKN-28 cells, as monitored by an MTT colorimetric assay. From the various compounds that were evaluated for their activity against HSV-1 and HSV-2, brivudine (BVDU) emerged as the most effective. When the 50% effective concentration (EC50) values of the antiherpes agents in MKN-28 cells were compared with those in human embryo lung MRC-5 cells, all compounds, except for BVDU, showed higher EC50 values in MKN-28 cells. For BVDU the EC50 values in MKN-28 cells were 0.8 (HSV-1) and 0.036 (HSV-2) times the EC50 values in MRC-5 cells. Thus BVDU was 27.5 times more active against HSV-2 in MKN-28 cells than in MRC-5 cells. The MKN-28 gastric cancer cells may be useful for the rapid screening of anti-HSV agents and, in particular, those that may be useful in therapy of gastrointestinal HSV infections in gastrointestinal herpetic infection.
Topics: Adenocarcinoma; Antiviral Agents; Bromodeoxyuridine; Colorimetry; Coloring Agents; Drug Evaluation, Preclinical; Herpesvirus 1, Human; Herpesvirus 2, Human; Humans; Stomach Neoplasms; Tetrazolium Salts; Thiazoles; Tumor Cells, Cultured; Virus Replication
PubMed: 8805100
DOI: 10.1111/j.1348-0421.1996.tb01080.x -
The British Journal of Ophthalmology Nov 1991The results of a randomised double-blind clinical trial of 0.1% bromovinyldeoxyuridine (BVDU) and 1% trifluorothymidine (TFT) in 60 patients with corneal dendritic... (Clinical Trial)
Clinical Trial Comparative Study Randomized Controlled Trial
The results of a randomised double-blind clinical trial of 0.1% bromovinyldeoxyuridine (BVDU) and 1% trifluorothymidine (TFT) in 60 patients with corneal dendritic ulceration are presented. There was no significant difference between BVDU and TFT in terms of numbers of ulcers healed (p = 0.61), mean healing time (p = 0.065), and cumulative healing rate (p = 0.058). No serious side effects were observed, though transient stinging was recorded in five patients receiving TFT and in three patients receiving BVDU. One patient in the group treated with TFT developed a punctate epitheliopathy.
Topics: Adolescent; Adult; Aged; Antiviral Agents; Bromodeoxyuridine; Child; Corneal Ulcer; Double-Blind Method; Female; Humans; Keratitis, Dendritic; Male; Middle Aged; Time Factors; Trifluridine; Wound Healing
PubMed: 1751456
DOI: 10.1136/bjo.75.11.649 -
Antimicrobial Agents and Chemotherapy Jul 1988The effect of 1-beta-D-arabinofuranosyl-E-5-(2-bromovinyl)uracil (BV-araU), a new antiviral drug, on Epstein-Barr virus (EBV) was studied and compared with those of... (Comparative Study)
Comparative Study
Comparison of two bromovinyl nucleoside analogs, 1-beta-D-arabinofuranosyl-E-5-(2-bromovinyl)uracil and E-5-(2-bromovinyl)-2'-deoxyuridine, with acyclovir in inhibition of Epstein-Barr virus replication.
The effect of 1-beta-D-arabinofuranosyl-E-5-(2-bromovinyl)uracil (BV-araU), a new antiviral drug, on Epstein-Barr virus (EBV) was studied and compared with those of E-5-(2-bromovinyl)-2'-deoxyuridine (BVdU) and acyclovir (ACV). BV-araU effectively inhibited EBV replication both in superinfected Raji cells and in virus producer P3HR-1(LS) cells, as determined by density gradient centrifugation, in situ cytohybridization with an EBV DNA probe, and cRNA-DNA hybridization. The 50% effective doses for viral DNA replication were 0.26, 0.06, and 0.3 microM for BV-araU, BVdU, and ACV, respectively. The relative efficacy on the basis of the in vitro therapeutic index was BVdU (6,500) greater than BV-araU (1,500) greater than ACV (850). Synthesis of EBV-induced polypeptides with molecular weights of 145,000 and 140,000 was inhibited by these drugs. Kinetic analysis of reversibility of inhibition of EBV DNA replication after removal of the drugs indicated that BV-araU, like BVdU, has a more prolonged inhibitory effect than ACV. These results indicate that the 2' OH group in the arabinosyl configuration of BV-araU results in marked reduction in anti-EBV activity while slightly diminishing cytotoxicity.
Topics: Acyclovir; Arabinofuranosyluracil; Bromodeoxyuridine; DNA, Viral; Dose-Response Relationship, Drug; Drug Combinations; Herpesvirus 4, Human; Peptide Biosynthesis; Uridine; Virus Replication
PubMed: 2847639
DOI: 10.1128/AAC.32.7.1068