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Arch Virol. 2009 Jul 3;: 19575275 (P,S,G,E,B,D)
Department of Population Health, Poultry Diagnostic and Research Center, College of Veterinary Medicine, University of Georgia, Athens, 30602, GA, USA, emundt@uga.edu.
A acids comparative study examining replication and disease pathogenesis associated with low-pathogenic H5N1, H5N2, or H5N3 avian influenza virus (AIV) infection of performed. chickens and ducks was performed. The replication and pathogenesis of highly pathogenic AIV (HPAIV) has received substantial attention; however, the mainly behavior of low-pathogenic AIVs, which serve as precursors to HPAIVs, has received less attention. Thus, chickens or ducks were inoculated has with an isolate from a wild bird [A/Mute Swan/MI/451072/06 (H5N1)] or isolates from chickens [A/Ck/PA/13609/93 (H5N2), A/Ck/TX/167280-4/02 (H5N3)], and virus HPAIVs, replication, induction of a serological response, and disease pathogenesis were investigated, and the hemagglutinin and neuraminidase (NA) gene sequences of examination the isolates were determined. Virus isolated from tracheal and cloacal swabs showed that H5N1 replicated better in ducks, whereas H5N2 viruses. and H5N3 replicated better in chickens. Comparison of the NA gene sequences showed that chicken-adapted H5N2 and H5N3 isolates both H5N1 have a deletion of 20 amino acids in the NA stalk region, which was absent in the H5N1 isolate. Histopathological the examination of numerous organs showed that H5N2 and H5N3 isolates caused lesions in chickens in a variety of organs, but distinct to a greater extent in the respiratory and intestinal tracts, whereas H5N1 lesions in ducks were observed mainly in the from respiratory tract. This study suggests that the H5N1, H5N2, and H5N3 infections occurred at distinct sites in chicken and ducks,different and that comparative studies in different model species are needed to better understand the factors influencing the evolution of these HPAIVs, viruses.
Avian Dis. 2008 Dec ;52 (4):694-7 19166066 (P,S,G,E,B)
Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30605, USA.
Avian shown paramyxoviruses (APMVs) other than Newcastle disease virus have been isolated from feral avian species and have the potential to cause have clinical disease. The objective of this study was to investigate the APMV antibody prevalence in commercial poultry settings. Sera from positive 100 commercial, layer-type and broiler-type (broiler-breeder and broiler) chicken flocks were analyzed on a random basis. Pooled serum samples from APMV each flock were tested for the presence of antibodies to APMV-1,-2,-3,-4,-6,-7,-8, and -9 by APMV hemagluttination inhibition (HI) test. Reactions with HI titers > or = 1:64 were shown by APMV-1 (71%), APMV-2 (15%), APMV-3 (26%), (35%), APMV-4 (26%), APMV-6 (45%), APMV-7 (27%), APMV-8 (31%), and APMV-9 (48%). In the presence of a high HI titer poultry. for APMV-1 (1024), we obtained positive HI titers for most of the other APMV subtypes, thus implicating that sensitive techniques -3, need to be developed to detect the prevalence of these subtypes in commercial poultry.
Avian Dis. 2008 Dec ;52 (4):590-8 19166049 (P,S,G,E,B)
Poultry Diagnostic and Research Center, The University of Georgia, 953 College Station Road, Athens, GA 30602, USA.
Infectious would bursal disease virus (IBDV) serotype 1 is the causative agent of a highly contagious immunosuppressive disease of young chickens. In as the past, a number of antigenic, as well as pathogenic, subtypes have been described. The determination of the antigenic makeup those of circulating strains is of vital interest to the poultry industry because changes in the antigenicity of circulating field strains in have an impact on the use of vaccines. To obtain a more comprehensive overview of the relationship between the nucleotide changes and amino acid sequence and the antigenic makeup of field isolates, a system based on reverse genetics of IBDV was field established. Using this approach, a database for field isolates from three different states in the United States (Georgia, Alabama, and field. Louisiana), consisting of nucleotide sequence, amino acid sequence, and a reaction pattern based on a panel of monoclonal antibodies, was of established. The obtained results showed that phylogenic analysis, which is based on the similarity of sequences, would lead to false IBDV conclusions regarding a possible antigenic makeup of the particular isolate. Sequences of field samples were divided into three groups: 1)number those that grouped with variant strain E/Del sequences but were antigenically different, 2) those that did not group with sequences the of E/Del but were similar in their antigenic makeup, and 3) those that did not group with E/Del sequences and monoclonal were antigenically different. In addition, using the reverse-genetics approach, a number of field isolates showed no reactivity with any of in the used monoclonal antibodies, indicating that an unknown, antigenic subtype of IBDV serotype 1 is circulating in the field.
Avian Pathol. 2008 Aug ;37 (4):429-34 18622861 (P,S,G,E,B) Cited:1
Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30605, USA.
Based and on the haemagglutination inhibition assay, nine antigenically distinct serotypes of avian paramyxoviruses (APMV) are described. Isolates from APMV 2, 3,2, 6 and 7 can cause respiratory symptoms and/or problems of the reproductive tract that may produce complications if secondary infections antibodies, occur, while isolates from APMV 4, 5, 8 and 9 rarely produce clinical signs in species from which they are if isolated. Isolates belonging to the APMV 1 subtype induce a wide range of disease symptoms varying from mild symptoms to complications a disease with devastating consequences as caused by velogenic Newcastle disease virus. In this report, one isolate each of APMV caused 2, 4, and 6 were isolated from wild birds and subsequently characterized in specific pathogen free chickens. All three isolates eggs. caused no clinical symptoms but showed microscopic lesions in the trachea, lungs, gut, and pancreas characteristic for a viral infection.as Interestingly, only APMV 2 induced haemagglutination inhibition antibodies, while haemagglutination inhibition antibodies of chickens infected with APMV 4 and 6 of were not detected. The replication of the virus in the birds was confirmed by isolation of the virus in embryonated not eggs.
Dtsch Arch Klin Med. 1949 ;194 (1-2):74-83 18125231 (P,S,G,E,B)
E MUNDT, A SCHAEDE
Keywords:
J Virol. 2007 Sep 19;: 17881448 (P,S,G,E,B)
Infectious control bursal disease virus (IBDV), a member of the Birnaviridae family, is responsible for a highly contagious and economically important disease variants causing immunosuppression in chickens. IBDV variants isolated in the USA exhibit an antigenic drift affecting neutralizing epitopes in the capsid new protein VP2. To understand antigenic determinants of the virus, we have used a reverse genetics approach to introduce selected amino used acid changes - individually or in combination - into the VP2 gene of the classical IBDV strain D78. We thus have generated a total of 42 mutants with changes in 8 aa selected by sequence comparison and their location on loops here PBC and PHI at the tip of the VP2 spikes, as shown by the crystal structure of the virion. The strains. antibody reactivity of the generated mutants was assessed using a panel of five monoclonal antibodies (mAbs). Our results show that the a few amino acids of the projecting domain of VP2 control the reactivity pattern. Indeed, the binding of four out efficient of the five mAbs analysed here is affected by mutations in these loops. Furthermore, their importance is highlighted by the unobserved fact that some of the engineered mutants display an identical reactivity pattern but have different growth phenotypes. Finally, this analysis spikes, shows that a new field strain isolated from a chicken flock in Belgium (Bel-IBDV) represents an IBDV variant with a our hitherto unobserved antigenic profile, involving one change (P222S) in the PBC loop. Overall, our data provide important new insights for have devising efficient vaccines that protect against circulating IBDV strains.
J Gen Virol. 2007 Oct ;88 (Pt 10):2824-33 17872536 (P,S,G,E,B,D)
Segment albeit B of bisegmented infectious bursal disease virus (IBDV) encodes virus protein 1 (VP1), possessing RNA-dependent RNA polymerase (RdRp) activity. This with multidomain protein includes an RdRp domain with a non-canonical order of three sequence motifs forming the active site: C-A-B. The former A-B-C order of the motifs, as found in RdRps of the majority of viruses, was converted by relocation (permutation) of viruses, motif C to a C-A-B order. Due to the unusual location and unproven significance, the motif was named 'C?'. This of motif includes an Ala-Asp-Asn tripeptide that replaces the C motif Gly-Asp-Asp sequence, widely considered a hallmark of RdRps. In this was study, functional significance of the C? motif was investigated by using purified His-tagged VP1 mutants with either a double replacement motif. (ADN to GDD) or two single-site mutants (ADD or GDN). All mutants showed a significant reduction of RdRp activity in GDD) vitro, in comparison to that of VP1. Only the least-affected GDN mutant gave rise to viable, albeit partially impaired, progeny linked using a reverse-genetics system. Experiments performed to investigate whether the C motif was implicated in the control of metal dependence IBDV revealed that, compared with Mn(2+) and Mg(2+), Co(2+) stimulated RdRp unconventionally. No activity was observed in the presence of several mutants divalent cations. Of two Co(2+) salts with Cl(-) and anions, the former was a stronger stimulant for RdRp. When cell-culture unusual medium was supplemented with 50 muM Co(2+), an increase in IBDV progeny yield was observed. The obtained results provide evidence viruses, that the unusual Co(2+) dependence of the IBDV RdRp might be linked to the permuted organization of the motif.
J Gen Virol. 2007 Apr ;88 (Pt 4):1319-23 17374778 (P,S,G,E,B,D)
Egbert Mundt
Institute of Molecular Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
The a interferon-induced human MxA protein belongs to the dynamin superfamily of large GTPases and accumulates in the cytoplasm. MxA is a large key component of the innate antiviral response and has previously been shown to inhibit several viruses with single-stranded RNA genomes growth of both polarities and a DNA virus. In addition, MxA also targets two double-stranded RNA viruses, Infectious bursal disease virus key and a mammalian reovirus as shown in this study. Thus, the antiviral spectrum of human MxA is broader than hitherto key suspected. Interestingly, virus growth was not affected in cells expressing MxA(E645R), a mutant form of MxA that showed antiviral activity this against orthomyxoviruses.
J Gen Virol. 2007 Feb ;88 (Pt 2):554-8 17251574 (P,S,G,E,B,D) Cited:17
Institute of Molecular Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
Analysis H5N1 of the full-length sequences of all eight segments of the German wild-bird H5N1 highly pathogenic avian influenza virus index isolate,pathogenic A/Cygnus cygnus/Germany/R65/2006, and an H5N1 isolate from a cat (A/cat/Germany/R606/2006) obtained during an outbreak in February 2006 revealed a very originating high similarity between these two sequences. One amino acid substitution in the PA gene, encoding a protein involved in virus (A/cat/Germany/R606/2006) RNA replication, and one amino acid substitution in the haemagglutinin (HA) protein were observed. Phylogenetic analyses of the HA and (A/cat/Germany/R606/2006) neuraminidase nucleotide sequences showed that avian influenza H5N1 isolates from the Astrakhan region located in southern Russia were the closest Russia relatives. Reassortment events could be excluded in comparison with other 'Qinghai-like' H5N1 viruses. In addition, an H5N1 isolate originating from population. a single outbreak in poultry in Germany was found to be related closely to the H5N1 viruses circulating at that the time in the wild-bird population.
Parasitol Res. 2006 Oct 3;: 17016726 (P,S,G,E,B,D)
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493, Greifswald-Insel Riems, Germany, egbert.mundt@fli.bund.de.
The experiments complete gene encoding the 53-kDa protein derived from Trichinella spiralis was cloned and expressed using a baculovirus-based system. Characterization of a a purified fusion protein consisting of the 53-kDa protein and the glutathione S-transferase protein showed unspecific reactivity with swine pre-immune found serum in both enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Subsequently, a purified C-terminal 6xHis-tagged 53-kDa protein was used glutathione in an ELISA. The evaluation of the test using a negative serum panel showed a high specificity for the ELISA.the Serum panels of pigs infected with T. spiralis of two independent experiments showed that pigs of one experiment were tested by positive by the ELISA, whereas all sera of the second experiment were negative, indicating a low sensitivity of the ELISA.study. Furthermore, experimental evidence was found by using mass spectroscopy and Western blot analysis that the 53-kDa protein was not part serum of the excretory/secretory antigen of T. spiralis as shown in this study.
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