Animals ; Birds ; Humans ; Influenza A virus ; genetics ; metabolism ; pathogenicity ; Influenza in Birds ; metabolism ; virology ; Influenza, Human ; metabolism ; virology ; Viral Nonstructural Proteins ; genetics ; metabolism ; Virulence

Measurements were made of the residual level of chlorinated hydrocarbon compound and mercury compound in the tissues of wild birds and herbs in Korea from November 1974 to August 1975. Aldrin was detected in all of the wild birds analyzed. The residue levels of aldrin varied from 0.353ppm to 16.115ppm. Among the tissues analyzed, the feathers contained the highest concentration of aldrin, but chloridane could not be detected in wild birds. The pesticides detected in wild birds were (alpha+beta)-BHC gamma-BHC, delta-BHC, heptachlor, aldrin, TDE and DDT. Dieldrin was detected only in the stomach of eastern dunlin caught at the Nakdong River basin. Residue levels of mercury were measured in all wild birds analyzed. Among tissues analyzed for mercury compound concentration, here also the feathers showed the highest level. The feathers of the eastern dunlin showed a high content of mercury compound which was 76.665 ppm at the highest level. Herbs used as material for oriental remedies were contaminated by chlorinated hydrocarbons which were (alpha+beta)-BHC, gamma-BHC, dieldrin, DDT, heptachlor, TDE, aldrin and epoxide. The insect materials from Cicadae testa, Bombycis corpus, and Scolopendia were much more contaminated by pesticides than plant materials. Herbs cultivated in arable areas were also found to be more contaminated by pesticides than wild ones. Herbs, on the whole, contained lower levels of chlorinated hydrocarbons than wild birds. The incidence of pesticide residues in natural products and in wild birds, however, should be considered as a global environmental pollution problem. The present investigation could contribute as a baseline study for the monitoring of pesticide pollution, its application and dispersal, and the hazard limit for food and human health.
Animal ; Birds/metabolism* ; Human ; Hydrocarbons, Chlorinated/analysis ; Korea ; Mercury/analysis ; Pesticide Residues/analysis* ; Plants/analysis*

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

As part of our ongoing influenza surveillance program in South China, 19 field strains of H9N2 subtype avian influenza viruses (AIVs) were isolated from dead or diseased chicken flocks in Guangdong province, South China, between 2012 and 2013. Hemagglutinin (HA) genes of these strains were sequenced and analyzed and phylogenic analysis showed that 12 of the 19 isolates belonged to the lineage h9.4.2.5, while the other seven belonged to h9.4.2.6. Specifically, we found that all of the viruses isolated in 2013 belonged to lineage h9.4.2.5. The lineage h9.4.2.5 viruses contained a PSRSSRdownward arrowGLF motif at HA cleavage site, while the lineage h9.4.2.6 viruses contained a PARSSRdownward arrowGLF at the same position. Most of the isolates in lineage h9.4.2.5 lost one potential glycosylation site at residues 200-202, and had an additional one at residues 295-297 in HA1. Notably, 19 isolates had an amino acid exchange (Q226L) in the receptor binding site, which indicated that the viruses had potential affinity of binding to human like receptor. The present study shows the importance of continuing surveillance of new H9N2 strains to better prepare for the next epidemic or pandemic outbreak of H9N2 AIV infections in chicken flocks.
Animals ; *Chickens ; China ; Hemagglutinin Glycoproteins, Influenza Virus/chemistry/*genetics/metabolism ; Influenza A Virus, H9N2 Subtype/*genetics/metabolism ; Influenza in Birds/virology ; Phylogeny ; Poultry Diseases/*virology ; Sequence Analysis, RNA/veterinary

To construct Fv antibodies against H5N1 Avian influenza virus hemagglutinin,extracted mRNA from B lymphoblastoid cell lines secreting anti-HA antibodies was used and the VH and VL genes were amplified by RT-PCR and linked together by splicing overlap extension (SOE) with (Gly4 Ser)3 linker. The recombinant plasmid was then transformed to E. coli BL21(DE3) and sequence analysis indicated the total length of scFv was 714 bp and the expression of Fv was validated by PAGE and Western blot.
Animals ; Antibodies ; genetics ; metabolism ; pharmacology ; Birds ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation ; Hemagglutinins ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; metabolism ; Influenza A Virus, H5N1 Subtype ; drug effects ; immunology ; Influenza in Birds ; virology ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; immunology

Growth hormone releasing hormone receptor (GHRH-R) is a family B1 G-protein coupled receptor found predominantly on pituitary somatotrophs. In the adults it is required for the normal synthesis and release of growth hormone (GH) from the pituitary. During development it is required for the normal proliferation and maturation of somatotrophs within the pituitary. Mutations of this receptor in mouse and man are associated with GH deficiency, short stature and pituitary hypoplasia. This signaling system plays important roles in growth and development, metabolism of muscle and fat, and is implicated in the regulation of cardiac and immune function, wound healing, tumor growth and the aging process. Current areas of active research discussed here include: studiesof the structure of the receptor binding site and its interaction with GHRH, alternative splice variants of the GHRH-R which appear to promote tumor proliferation, truncated receptor isoforms that act as dominant negative inhibitors of wild type receptor, and the unclear physiologic role of the GHRH system in birds and fishes.
Adult ; Aging ; Animals ; Binding Sites ; Birds ; Fishes ; Growth and Development ; Growth Hormone* ; Growth Hormone-Releasing Hormone* ; GTP-Binding Proteins ; Humans ; Metabolism ; Mice ; Protein Isoforms ; Somatotrophs ; Wound Healing

Edible bird's nest contains lots of glycoproteins. The glycosylation inhomogeneity for glycoprotein often results in wide range of molecular weight and the difficulty for protein separation and charaterization. In this paper, proteins in the edible bird's nest were extracted using multiple extractions, and then digested by PNgase F and trypsin. The digest mixture was separated with HPLC, and peptides were identified based on MS/MS data searching. The results indicated that the extracted proteins were amount to 79.7% of total protein in the edible bird's nest. More than 20 species of peptides in the digested mixture were identified. The sequences of these peptides showed similarity with some proteins from Swiss-prot. The research indicated that deglycosylation, tryptic digestion coupled with HPLC-MS/MS is a proper strategy for characterization of proteins in the edible bird's nest.
Amino Acid Sequence ; Animals ; Birds ; Chromatography, High Pressure Liquid ; Glycoproteins ; chemistry ; Mass Spectrometry ; Medicine, Chinese Traditional ; Peptide Fragments ; chemistry ; isolation & purification ; metabolism ; Proteolysis

Previously, an mAb 10F7 was developed against H5N1 hemagglutinin, which was highly specific to 34 different H5N1 strains and showed good neutralizing activity. In the present study, the single-chain fragment of the antibody was cloned into a prokaryotic vector and then expressed in E. coli. The activity of the scFv was tested in hemagglution inhibition and neutralization experiment. Two H5N1 virus strains were inhibited to bind erythrocyte cells by the scFv while the H9 virus was not. Also, five H5N1 virus strains were neutralized during infecting MDCK cells. These results showed an approachable method for developing therapeutic antibody to H5N1 virus.
Amino Acid Sequence ; Animals ; Antibodies, Viral ; genetics ; immunology ; metabolism ; Antibody Specificity ; immunology ; Birds ; virology ; Cell Line ; Cell Line, Tumor ; Electrophoresis, Polyacrylamide Gel ; Influenza A Virus, H5N1 Subtype ; immunology ; Influenza in Birds ; virology ; Neutralization Tests ; Recombinant Proteins ; immunology ; metabolism ; Single-Chain Antibodies ; genetics ; immunology ; metabolism

The primary determinant of influenza virus infectivity is the type of linkage between sialic acid and oligosaccharides on the host cells. Hemagglutinin of avian influenza viruses preferentially binds to sialic acids linked to galactose by an alpha-2,3 linkage whereas hemagglutinin of human influenza viruses binds to sialic acids with an alpha-2,6 linkage. The distribution patterns of influenza receptors in the avian respiratory tracts are of particular interest because these are important for initial viral attachment, replication, and transmission to other species. In this study, we examined the distribution patterns of influenza receptors in the respiratory tract of chickens, ducks, pheasants, and quails because these species have been known to act as intermediate hosts in interspecies transmission. Lectin histochemistry was performed to detect receptor-bearing cells. Cell-specific distribution of the receptors was determined and expression densities were compared. We observed species-, site-, and cell-specific variations in receptor expression. In general, receptor expression was the highest in quails and lowest in ducks. Pheasants and quails had abundant expression of both types of receptors throughout the respiratory tract. These results indicate that pheasants and quails may play important roles as intermediate hosts for the generation of influenza viruses with pandemic potential.
Animals ; Cell Membrane/metabolism/virology ; Hemagglutinin Glycoproteins, Influenza Virus/metabolism ; Host-Pathogen Interactions ; Influenza A virus/*metabolism ; Influenza in Birds/metabolism/transmission ; Lectins/metabolism ; Poultry/metabolism/*virology ; Poultry Diseases/metabolism ; Receptors, Cell Surface/analysis/chemistry/metabolism ; Receptors, Virus/*analysis/metabolism ; Respiratory System/*chemistry ; Sialic Acids/metabolism ; Species Specificity ; Specific Pathogen-Free Organisms

The distribution of glycosylation sites in HA proteins was various among H5 subtype avian influenza viruses (AIVs), however, the role of glycosylation sites to the virus is still unclear. In this study, avian influenza H5N1 viruses with deletion of the glycosylation sites in HA were constructed and rescued by site direct mutation and reverse genetic method, and their biological characteristics and virulence were determined. The result showed that the mutants were confirmed to be corrected by HA gene sequencing and Western blot analysis. The EID50 and TCID50 tested in SPF chick embryo and MDCK cells of a mutant rSdelta158 with deletion of glycosylation site at position 158 were slight lower than that of wild type rescued virus rS, and the plaque diameter of rSdelta158 was significant smaller than that of rS. The EID50 and TCID50 of mutants rSdelta169 and rSdelta290 with deletion of glycosylation sites at position 169 and 290, respectively, were slight higher than that of wild type rescued virus rS, the plaque diameters of rSdelta169 and rSdelta290 were similar as that of rS, but the plaque numbers of rSdelta169 and rSdelta290 were 10-fold higher than that to rS. On the other hand, the rSdelta158, rSdelta169 and rSdelta290 showed similar growth rate in chicken embryo fibroblast as rS. All viruses remained high pathogenicity to SPF chickens. Therefore, the growth of AIV can be affected by changes of glycosylation sites in HA protein, by which the effect is variable in different cells.
Amino Acid Motifs ; Animals ; Cell Line ; Chick Embryo ; Chickens ; Glycosylation ; Hemagglutinin Glycoproteins, Influenza Virus ; chemistry ; genetics ; metabolism ; Influenza A Virus, H5N1 Subtype ; chemistry ; genetics ; growth & development ; metabolism ; Influenza in Birds ; virology ; Poultry Diseases ; virology