Prion proteins (PrPs) are infectious pathogens that cause a group of invariably fatal, neurodegenerative diseases, including Creutzfeldt-Jakob disease, by means of an entirely novel mechanism. They are produced by various species, including reptile, rodent, ruminant and mammals, during normal metabolic processes, but they can be slowly changed into pathogenic isoforms upon contact with other infectious PrP isoforms. This transmission can occur across species barriers. In the present study, phylogram for each PrP sequence was generated by PAUP* 4.0 program using Neighbor-Joining method with 1,000 times bootstrapping process for the phylogenetic analysis. The molecular dynamics (MD) simulations were performed by the SANDER module in the AMBER 7 package using Amber 99 force field. All the simulation process was conducted in the IBM p690 Supercomputing System in Korea Institute of Science and Technology Information. To reduce the calculation time, we used the Generalized Born (GB) model. We compared the sequences and structural characteristics of normal and pathogenic (E200K) human PrPs with those of other reptile, rodent, ruminant and mammalian PrPs. Phylogenetic analysis revealed that, although the turtle PrP sequence is the most distinct of the PrPs analyzed, it nonetheless retains five conserved secondary structural elements that are similar to those found in the mammalian PrPs, suggesting that these elements have important functions in vivo. The RMS deviation between the normal and E200K human PrPs was larger than that between the normal human and bovine PrPs, and all of the beta-sheet structures in human E200K PrP were very stable during MD simulations.
Animals ; Cattle ; *Computational Biology ; Humans ; Phylogeny ; Prions/*chemistry/classification/*genetics/isolation & purification ; Reptiles/metabolism ; Rodentia/metabolism ; Ruminants/metabolism ; Sequence Analysis, Protein ; Species Specificity

The nucleotide sequences of P gene from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The P gene is 1,655 nucleotides long with two overlapping open reading frames (ORFs). The first ORF is 1530 nucleotides long and would produce P protein of 509 amino acid residues. The second ORF is 534 nucleotides long and would produce C protein of 177 amino acid residues. The first ORF produces a second mRNA transcript of 897 nucleotides long with an extra G nucleotide at position 751. Translation from this mRNA would produce V protein of 298 amino acid residues. The nucleotide and deduced amino acid sequence were compared with the homologous region of other PPRV isolates. At the amino acid level, the "China/Tib/Gej/07-30" shares homology of 86.10%-97.3%, 84.3%-94.9%, and 82.9%-96.3% for P, C, and V proteins respectively. Several sequence motifs in the P genes were identified on the basis of conservation in the PPRVs and the morbilliviruses.
Amino Acid Sequence ; Animals ; China ; Female ; Goat Diseases ; virology ; Goats ; Molecular Sequence Data ; Peste-des-Petits-Ruminants ; veterinary ; virology ; Peste-des-petits-ruminants virus ; chemistry ; genetics ; isolation & purification ; metabolism ; Phosphoproteins ; chemistry ; genetics ; metabolism ; Sequence Analysis ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics ; metabolism

The purpose of the study is to construct recombinant goat pox virus (GPV) expressing Peste des petits ruminants virus (PPRV) H protein, and to evaluate the immunization effect. Recombinant GPV containing PPRV H gene (rGPV-PPRV-H) was selected and purified by gpt and eGFP utilizing plaque purification, and the final selected recombinant GPV was proved to be purified by PCR. Immunofluorescence and Western blotting showed that the recombinant virus could express H protein of PPRV while infecting lamb testis cells. Six goats were immunized with 2 x 10(6) PFU rGPV-PPRV-H through intradermal injection, and were immunized for the second time at 28 days with the same dose recombinant virus after first immunization. Serum was collected after immunization, and was analyzed for the neutralization antibodies. 21 days after first immunization, the neutralization antibodies of GPV were 40, 80, > or = 80, > or = 80, 40, > or = 80 in turn, and neutralization antibodies of PPRV were 80, 80, 80, 80, 40, 40, 10 in turn; 14 days after second immunization, the neutralization antibodies of GPV were all > or = 80, and the neutralization antibodies of PPRV were > 80, 80, > 80, 80, 80 and 40 in turn. This study established a foundation for the industrialization of the PPRV recombinant GPV vaccine.
Animals ; Capripoxvirus ; genetics ; immunology ; Goat Diseases ; immunology ; prevention & control ; virology ; Goats ; Hemagglutinins, Viral ; genetics ; immunology ; metabolism ; Peste-des-Petits-Ruminants ; immunology ; prevention & control ; Peste-des-petits-ruminants virus ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Vaccines, Combined ; immunology ; Vaccines, Synthetic ; immunology ; Viral Vaccines ; immunology

OBJECTIVETo investigate the cardioprotective effects of the peptides from the velvet antler (VAP) of Cervus nippon on acute ischemic myocardial injury in rats and its underlying mechanisms.

METHODThe model rats of myocardial ischemia injury (MI) was produced by ligating the left anterior descending (LAD) branch of the coronary artery of rats. The rats were divided randomly into six groups: sham-operated group; ischemic myocardial injury (MI) model group; diltiazem hydrochloride 1 mg x kg(-1) group; VAP 30 mg x kg(-1) group; VAP 60 mg x kg(-1) group; VAP 120 mg x kg(-1) group. VAP or diltiazem hydrochloride (calcium-antagonist) was injected into the tail vein of each of the experimental rats 12 h and 23 h respectively after left anterior descending (LAD) occlusion. The same volume of saline was administered into the sham-operated group and MI model group. The ischemic size, ST elevation, serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate transaminase (AST), superoxide dismutase (SOD), malondialdehyde (MDA), and levels of SOD, MDA in myocardial tissue were measured in the rats at 24 h after LAD occlusion.

RESULTThe ischemic risk area ratio was 34.52 +/- 8.69% in the MI control group. Treatment of VAP at doses of 60 and 120 mg x kg(-1) resulted in reductions in the infarct size of (21.67 +/- 5.19)% and (19.56 +/- 9.15)% (P < 0.05), respectively. Treatment with VAP at doses of 30, 60 and 120 mg x kg(-1) significantly decreased ST elevation (0.28 +/- 0.03, 0.21 +/- 0.03, 0.16 +/- 0.02, 0.15 +/- 0.02) mV (all P < 0.01), compared with the MI control rats (0.41 +/- 0.05) mV. VAP also significantly decreased levels of CK, LDH, AST, MDA in serum and myocardial tissue, increase SOD activity in serum and myocardial tissue in MI rats.

CONCLUSIONVAP exerts significant cardioprotective effects against acute ischemic myocardial injury in rats, likely through its antioxidant and anti-lipid peroxidation properties.

Acute Disease ; therapy ; Animals ; Antlers ; chemistry ; metabolism ; Cardiotonic Agents ; therapeutic use ; Disease Models, Animal ; Humans ; Male ; Myocardial Reperfusion Injury ; drug therapy ; Peptides ; therapeutic use ; Random Allocation ; Rats ; Rats, Wistar ; Ruminants ; metabolism