scholarly journals Evolution of the Sabin Strain of Type 3 Poliovirus in an Immunodeficient Patient during the Entire 637-Day Period of Virus Excretion

2000 ◽  
Vol 74 (7) ◽  
pp. 3001-3010 ◽  
Author(s):  
Javier Martín ◽  
Glynis Dunn ◽  
Robin Hull ◽  
Varsha Patel ◽  
Philip D. Minor
Keyword(s):  
Temperature Sensitive ◽  
Wild Type ◽  
Human Gut ◽  
Wild Poliovirus ◽  
Antigenic Properties ◽  
Virus Excretion ◽  
Main Lineage ◽  
Type 3

ABSTRACT A 20-year-old female hypogammaglobulinemic patient received monotypic Sabin 3 vaccine in 1962. The patient excreted type 3 poliovirus for a period of 637 days without developing any symptoms of poliomyelitis, after which excretion appeared to have ceased spontaneously. The evolution of Sabin 3 throughout the entire period of virus excretion was studied by characterization of seven sequential isolates from the patient. The isolates were analyzed in terms of their antigenic properties, virulence, sensitivity for growth at high temperatures, and differences in nucleotide sequence from the Sabin type 3 vaccine. The isolates followed a main lineage of evolution with a rate of nucleotide substitution that was very similar to that estimated for wild-type poliovirus during person-to-person transmission. There was a delay in the appearance of antigenic variants compared to sequential type 3 isolates from healthy vaccines, which could be one of the possible explanations for the long-term excretion of virus from the patient. The distribution of mutations in the isolates identified regions of the virus possibly involved in adaptation for growth in the human gut and virus persistence. None of the isolates showed a full reversion of the attenuated and temperature-sensitive phenotypes of Sabin 3. Information of this sort will help in the assessment of the risk of spread of virulent polioviruses from long-term excretors and in the design of therapies to stop long-term excretion. This will make an important contribution to the decision-making process on when to stop vaccination once wild poliovirus has been eradicated.

scholarly journals Antigenic and Molecular Characterization of Wild Type 1 Poliovirus Causing Outbreaks of Poliomyelitis in Albania and Neighboring Countries in 1996

1998 ◽  
Vol 36 (7) ◽  
pp. 1912-1918 ◽  
Author(s):  
L. Fiore ◽  
D. Genovese ◽  
E. Diamanti ◽  
S. Catone ◽  
B. Ridolfi ◽  
...  
Keyword(s):  
Western Europe ◽  
Antigenic Structure ◽  
Wild Type ◽  
Wild Poliovirus ◽  
The Past ◽  
The World ◽  
Large Outbreak ◽  
Human Sera

Mass vaccination has led poliomyelitis to become a rare disease in a large part of the world, including Western Europe. However, in the past 20 years wild polioviruses imported from countries where polio is endemic have been responsible for outbreaks in otherwise polio-free European countries. We report on the characterization of poliovirus isolates from a large outbreak of poliomyelitis that occurred in Albania in 1996 and that also spread to the neighboring countries of Yugoslavia and Greece. The epidemics involved 145 subjects, mostly young adults, and caused persisting paralysis in 87 individuals and 16 deaths. The agent responsible for the outbreak was isolated from 74 patients and was identified as wild type 1 poliovirus by both immunological and molecular methods. Sequence analysis of the genome demonstrated the involvement of a single virus strain throughout the epidemics, and genotyping analysis showed 95% homology of the strain with a wild type 1 poliovirus strain isolated in Pakistan in 1995. Neutralization assays with both human sera and monoclonal antibodies were performed to analyze the antigenic structure of the epidemic strain, suggesting its peculiar antigenic characteristics. The presented data underline the current risks of outbreaks due to imported wild poliovirus and emphasize the need to improve vaccination efforts and also the need to implement surveillance in countries free of indigenous wild poliovirus.

ucFabV Requires Functional Reductase Activity to Confer Reduced Triclosan Susceptibility in Escherichia coli

2015 ◽  
Vol 25 (6) ◽  
pp. 394-402 ◽  
Author(s):  
Taylor L. Fischer ◽  
Robert J. White ◽  
Katherine F.K. Mares ◽  
Devin E. Molnau ◽  
Justin J. Donato
Keyword(s):  
Escherichia Coli ◽  
Reductase Activity ◽  
Acid Synthesis ◽  
Temperature Sensitive ◽  
Wild Type ◽  
E Coli ◽  
Enoyl Acp Reductase ◽  
Selection For

<b><i>Background/Aims:</i></b> We previously identified the Triclo1 fosmid in a functional metagenomic selection for clones that increased triclosan tolerance in <i>Escherichia coli</i>. The active enzyme encoded by Triclo1 is ucFabV. Although ucFabV is homologous to FabV from other organisms, ucFabV contains substitutions at key positions that would predict differences in substrate binding. Therefore, a detailed characterization of ucFabV was conducted to link its biochemical activity to its ability to confer reduced triclosan sensitivity. <b><i>Methods:</i></b> ucFabV and a catalytic mutant were purified and used to reduce crotonoyl-CoA in vitro. The mutant and wild-type enzymes were introduced into <i>E. coli</i>, and their ability to confer triclosan tolerance as well as suppress a temperature-sensitive mutant of FabI were measured. <b><i>Results:</i></b> Purified ucFabV, but not the mutant, reduced crotonoyl-CoA in vitro. The wild-type enzyme confers increased triclosan tolerance when introduced into <i>E. coli</i>, whereas the mutant remained susceptible to triclosan<i>. </i>Additionally, wild-type ucFabV, but not the mutant, functionally replaced FabI within living cells. <b><i>Conclusion:</i></b> ucFabV confers increased tolerance through its function as an enoyl-ACP reductase. Furthermore, ucFabV is capable of restoring viability in the presence of compromised FabI, suggesting ucFabV is likely facilitating an alternate step within fatty acid synthesis, bypassing FabI inhibition.

scholarly journals CHARACTERIZATION OF DOMINANT RESISTANCE TO COBALT CHLORIDE IN DICTYOSTELIUM DISCOIDEUM AND ITS USE IN PARASEXUAL GENETIC ANALYSIS

Genetics ◽  
1978 ◽  
Vol 90 (1) ◽  
pp. 37-47
Author(s):  
Keith L Williams
Keyword(s):  
Dictyostelium Discoideum ◽  
Cobalt Chloride ◽  
Resistance Mutation ◽  
Differential Sensitivity ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Cobaltous Chloride ◽  
Chloride Resistance ◽  
Resistant Strains

ABSTRACT Strains of Dictyostelium discoideum resistant to cobaltous chloride have been isolated at a frequency of approximately 10-6. The resistant strains have one of three phenotypes, recessive to wild type, dominant to wild type and dominant to wild type but requiring the presence of cobaltous chloride to maintain resistance. Strains carrying a dominant cobaltous chloride resistance mutation and a recessive growth temperature-sensitive mutation can be mixed with wild-type haploid lines and then subjected to selection so that only diploid lines survive. Differential sensitivity to cycloheximide has also been observed. Hypersensitivity to cycloheximide in combination with dominant cobaltous chloride resistance provides a means of selecting diploids without the use of temperature-sensitive mutations.

scholarly journals Cycloheximide-resistant temperature-sensitive lethal mutations of Saccharomyces cerevisiae.

Genetics ◽  
1988 ◽  
Vol 119 (2) ◽  
pp. 303-315
Author(s):  
J H McCusker ◽  
J E Haber
Keyword(s):  
Inhibitory Concentration ◽  
Temperature Sensitive ◽  
Nonpermissive Temperature ◽  
Wild Type ◽  
Lethal Mutations ◽  
New Methods ◽  
Similar Temperature ◽  
Complementation Groups ◽  
Wild Type Yeast

Abstract We describe the isolation and preliminary characterization of a set of pleiotropic mutations resistant to the minimum inhibitory concentration of cycloheximide and screened for ts (temperature-sensitive) growth. These mutations fall into 22 complementation groups of cycloheximide resistant ts lethal mutations (crl). None of the crl mutations appears to be allelic with previously isolated mutations. Fifteen of the CRL loci have been mapped. At the nonpermissive temperature (37 degrees), these mutants arrest late in the cell cycle after several cell divisions. Half of these mutants are also unable to grow at very low temperatures (5 degrees). Although mutants from all of the 22 complementation groups exhibit similar temperature-sensitive phenotypes, an extragenic suppressor of the ts lethality of crl3 does not relieve the ts lethality of most other crl mutants. A second suppressor mutation allows crl10, crl12, and crl14 to grow at 37 degrees but does not suppress the ts lethality of the remaining crl mutants. We also describe two new methods for the enrichment of auxotrophic mutations from a wild-type yeast strain.

scholarly journals CDC37 is required for p60v-src activity in yeast.

1996 ◽  
Vol 7 (9) ◽  
pp. 1405-1417 ◽  
Author(s):  
B Dey ◽  
J J Lightbody ◽  
F Boschelli
Keyword(s):  
Protein Tyrosine Kinase ◽  
Wild Type Strain ◽  
Active Form ◽  
Biologically Active ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Wild Type ◽  
Isolation And Characterization ◽  
Genes Encoding

Mutations in genes encoding the molecular chaperones Hsp90 and Ydj1p suppress the toxicity of the protein tyrosine kinase p60v-src in yeast by reducing its levels or its kinase activity. We describe isolation and characterization of novel p60v-src-resistant, temperature-sensitive cdc37 mutants, cdc37-34 and cdc37-17, which produce less p60v-src than the parental wild-type strain at 23 degrees C. However, p60v-src levels are not low enough to account for the resistance of these strains. Asynchronously growing cdc37-34 and cdc37-17 mutants arrest in G1 and G2/M when shifted from permissive temperatures (23 degrees C) to the restrictive temperature (37 degrees C), but hydroxyurea-synchronized cdc37-34 and cdc37-17 mutants arrest in G2/M when released from the hydroxyurea block and shifted from 23 to 37 degrees C. The previously described temperature-sensitive cdc37-1 mutant is p60v-src-sensitive and produces wild-type amounts of p60v-src at permissive temperatures but becomes p60v-src-resistant at its restrictive temperature, 38 degrees C. In all three cdc37 mutants, inactivation of Cdc37p by incubation at 38 degrees C reduces p60v-src-dependent tyrosine phosphorylation of yeast proteins to low or undetectable levels. Also, p60v-src levels are enriched in urea-solubilized extracts and depleted in detergent-solubilized extracts of all three cdc37 mutants prepared from cells incubated at the restrictive temperature. These results suggest that Cdc37p is required for maintenance of p60v-src in a soluble, biologically active form.

scholarly journals Isolation of epidemic poliovirus from sewage during the 1992–3 type 3 outbreak in the Netherlands

1995 ◽  
Vol 114 (3) ◽  
pp. 481-491 ◽  
Author(s):  
H. G. A. M. van der Avoort ◽  
J. H. J. Reimerink ◽  
A. Ras ◽  
M. N. Mulders ◽  
A. M. van Loon
Keyword(s):  
The Netherlands ◽  
Oral Poliovirus Vaccine ◽  
Wild Type ◽  
Risk Area ◽  
Poliovirus Type ◽  
Wild Poliovirus ◽  
Home Village ◽  
A Site ◽  
Religious Reasons ◽  
Type 3

SUMMARYTo examine the extent of wild poliovirus circulation during the 1992–3 epidemic in the Netherlands caused by poliovirus type 3, 269 samples from sewage pipelines at 120 locations were examined for the presence of poliovirus. The epidemic virus strain was found in 23 samples, all from locations inside the risk area which contained communities that refuse vaccination for religious reasons. By sewage investigation, the wildtype virus was shown to be present in the early phase of the epidemic at two locations, one week before patients were reported from that area. The wild type 3 poliovirus was also detected retrospectively in a river water sample collected for other reasons three weeks before notification of the first poliomyelitis case, at a site a few kilometres upstream the home village of this patient. Oral poliovirus vaccine (OPV) virus was found at 28 locations inside or at the border of the risk area. Trivalent OPV was offered to unvaccinated or incompletely-vaccinated persons living in this region as part of the measures to control the epidemic.

scholarly journals CHARACTERIZATION OF TEMPERATURE-SENSITIVE, FERTILIZATION-DEFECTIVE MUTANTS OF THE NEMATODE CAENORHABDITIS ELEGANS

Genetics ◽  
1978 ◽  
Vol 88 (2) ◽  
pp. 285-303 ◽  
Author(s):  
Samuel Ward ◽  
Johji Miwa
Keyword(s):  
Caenorhabditis Elegans ◽  
Sensitive Period ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Wild Type ◽  
Normal Numbers ◽  
Primary Defect ◽  
Isolation And Characterization ◽  
Pass Through

ABSTRACT The isolation and characterization of three Caenorhabditis elegans temperature-sensitive mutants that are defective at fertilization are described. All three are alleles of the gene fer-1. At the restrictive temperature of 25°, mutant hermaphrodites make sperm and oocytes in normal numbers. No oocytes are fertilized, although they pass through the spermatheca and uterus normally. The oocytes can be fertilized by sperm transferred by wild-type males, indicating that the mutant defect is in the sperm. The temperature-sensitive period for the mutants coincides with spermatogenesis. Sperm made by mutants at 25° cannot be distinguished from wild-type sperm by light microscopy. The sperm do contact oocytes in mutant hermaphrodites, but do not fertilize. Mutant sperm appear to be nonmotile. Mutant males are also sterile when grown at 25°. They transfer normal numbers of sperm to hermaphrodites at mating, but these sperm fail to migrate to the spermatheca and are infertile. The phenotype of these mutants is consistent with a primary defect in sperm motility, but the cause of this defect is not known.

Molecular characterization of a wild poliovirus type 3 epidemic in The Netherlands (1992 and 1993).

1995 ◽  
Vol 33 (12) ◽  
pp. 3252-3256 ◽  
Author(s):  
M N Mulders ◽  
A M van Loon ◽  
H G van der Avoort ◽  
J H Reimerink ◽  
A Ras ◽  
...  
Keyword(s):  
The Netherlands ◽  
Molecular Characterization ◽  
Poliovirus Type ◽  
Wild Poliovirus ◽  
Type 3

scholarly journals Natural Genetic Exchanges between Vaccine and Wild Poliovirus Strains in Humans

2000 ◽  
Vol 74 (18) ◽  
pp. 8434-8443 ◽  
Author(s):  
Sophie Guillot ◽  
Valérie Caro ◽  
Nancy Cuervo ◽  
Ekaterina Korotkova ◽  
Mariana Combiescu ◽  
...  
Keyword(s):  
Paralytic Poliomyelitis ◽  
Nucleotide Sequencing ◽  
Wild Type ◽  
Coding Region ◽  
Wild Poliovirus ◽  
Type 3 ◽  
Recombinant Genome ◽  
Natural Means

ABSTRACT In a previous study of poliovirus vaccine-derived strains isolated from patients with vaccine-associated paralytic poliomyelitis (VAPP) (9, 11), we reported that a high proportion (over 50%) of viruses had a recombinant genome. Most were intertypic vaccine/vaccine recombinants. However, some had restriction fragment length polymorphism (RFLP) profiles different from those of poliovirus vaccine strains. We demonstrate here that five such recombinants, of 88 VAPP strains examined, carried sequences of wild (nonvaccine) origin. To identify the parental wild donor of these sequences, we used RFLP profiles and nucleotide sequencing to look for similarity in the 3D polymerase-coding region of 61 wild, cocirculating poliovirus isolates (43 type 1, 16 type 2, and 2 type 3 isolates). In only one case was the donor identified, and it was a wild type 1 poliovirus. For the other four vaccine/wild recombinants, the wild parent could not be identified. The possibility that the wild sequences were of a non-poliovirus-enterovirus origin could not be excluded. Another vaccine/wild recombinant, isolated in Belarus from a VAPP case, indicated that the poliovirus vaccine/wild recombination is not an isolated phenomenon. We also found wild polioviruses (2 of 15) carrying vaccine-derived sequences in the 3′ moiety of their genome. All these results suggest that genetic exchanges with wild poliovirus and perhaps with nonpoliovirus enteroviruses, are also a natural means of evolution for poliovirus vaccine strains.

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