Amino Acid Sequence ; Genetic Structures ; Genetic Variation ; Genome, Viral ; Molecular Sequence Data ; SARS Virus ; genetics

PURPOSE: Although the mechanism of virus-induced, aspirin-exacerbated respiratory disease (AERD) is not known fully, direct activation of viral components through Toll-like receptor 3 (TLR3) has been suggested. TLR3 recognizes double-stranded RNA (dsRNA), and activates nuclear factor-kappaB and increases interferon-gamma, which signals other cells to induce airway inflammation in asthma. Considering the association of TLR3 in viral infections and AERD, we investigated whether promoter and non-synonymous variants of TLR3 were associated with AERD. METHODS: The three study groups, 203 with AERD, 254 with aspirin-tolerant asthma (ATA), and 274 normal healthy controls (NC) were recruited from Ajou University Hospital, Korea. Two polymorphisms, -299698G>T and 293391G>A [Leu412Phe], were genotyped using primer extension methods. RESULTS: Genetic associations were examined between two genetic polymorphisms of TLR3 (-299698G>T and 293391G>A [Leu412Phe]) in the three study groups. AERD patients that carried the GG genotype of 293391G>A showed a significantly lower frequency compared with ATA in both co-dominant (P=0.025) and dominant models (P=0.036). Similarly, in the minor allele frequency, the A allele was significantly higher (P=0.023) in AERD compared with ATA for this polymorphism. AERD patients who carried HT2 [GA] showed a significantly higher frequency than other haplotypes in co-dominant (P=0.02) and recessive (P=0.026) models. CONCLUSIONS: Our findings suggest that the -299698G>T and 293391G>A [Leu412Phe] polymorphisms of the TLR3 gene are associated with the AERD phenotype.
Alleles ; Asthma ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Inflammation ; Interferon-gamma ; Korea ; Phenotype ; Polymorphism, Genetic ; RNA, Double-Stranded ; Toll-Like Receptor 3 ; Toll-Like Receptors ; Viral Structures

According to the serological screening methods of antigen-antibody reaction such as ELISA, it has been known that the complete detection of viral infections of HBV, HCV, and HIV-1 viruses in the blood and blood related-products is not much reliable. Therefore, nucleic acid amplification testing methods (NAT) adopted to detect the small quantitative viral nucleic acids could support the basis of using and supplying the blood and its related products safely. This research work is basically designed to describe the simultaneous blood screening system by multiplex or duplex tests for detection of HBV, HCV, and HIV-1 viruses in the blood at one time with low price and labor. It is aimed at easy detection by using the conventional agarose gel electrophoresis. Thus, we tried to detect and identify the viral components in the blood sample according to their different size of PCR products. We decided a set of consensus sequences to recognize each viral DNA fragments after running the multiplex PCR in one tube. This was done by nested RT-PCR using two different RNA viral genomic templates followed by multiplex PCR with addition of viral DNA and their primers after purifying the viral genomic nucleic acids. Those specific primers could be used without any interference to amplify each viral genome in the blood samples. The sensitivities with different viral loads were evaluated on the agarose gel electrophoresis. Three different viral agents in the blood samples could be tested by this multiplex (RT)-PCR with three different primers.
Antigen-Antibody Reactions ; Consensus Sequence ; DNA, Viral ; Electrophoresis, Agar Gel ; Enzyme-Linked Immunosorbent Assay ; Genome, Viral ; HIV-1 ; Mass Screening ; Multiplex Polymerase Chain Reaction ; Nucleic Acid Amplification Techniques ; Nucleic Acids ; Polymerase Chain Reaction ; Quality Control* ; RNA ; Running ; Viral Load ; Viral Structures

OBJECTIVES: Vesicular stomatitis virus (VSV) is under development as an oncolytic virus due to its preferential replication in cancer cells and oncolytic activity, however the viral components responsible have not yet been determined. In this study the effects of VSV wild-type (wt) and M51R-mutant matrix proteins (M51R-mMP) on apoptosis, pyroptosis, necroptosis, and autophagy pathways, in an esophagus cancer cell line (KYSE-30) were investigated. METHODS: The KYSE-30 cells were transfected with pcDNA3.1 plasmids encoding wt or M51R-mMP, and apoptosis, pyroptosis, necroptosis, and autophagy were evaluated 48 and 72 hours after transfection. RESULTS: KYSE-30 cells transfected with VSV wt and M51R-mMPs significantly reduced cell viability to < 50% at 72 hours post-transfection. M51R-MP significantly increased the concentration of caspase-8 and caspase-9 at 48 and 72 hours post-transfection, respectively ( p < 0.05). In contrast, no significant changes were detected following transfection with the VSV wt plasmid. Moreover, VSV wt and M51R-mMP transfected cells did not change the expression of caspase-3. VSV wt and M51R-mMPs did not mMP change caspase-1 expression (a marker of pyroptosis) at 48 and 72 hours post-transfection. However, M51R-mMP and VSV wt transfected cells significantly increased RIP-1 (a marker of necroptosis) expression at 72 hours post-infection ( p < 0.05). Beclin-1, a biomarker of autophagy, was also induced by transfection with VSV wt or M51R-mMPs at 48 hours post-transfection. CONCLUSION: The results in this study indicated that VSV exerts oncolytic activity in KYSE-30 tumor cells through different cell death pathways, suggesting that M51R-mMP may potentially be used to enhance oncolysis.
Apoptosis ; Autophagy ; Carcinoma, Squamous Cell ; Caspase 3 ; Caspase 8 ; Caspase 9 ; Cell Death ; Cell Line ; Cell Survival ; Epithelial Cells ; Esophageal Neoplasms ; Oncolytic Viruses ; Plasmids ; Pyroptosis ; Transfection ; Vesicular Stomatitis ; Viral Structures

Adenoviruses are icosahedral virions containing double-stranded linen DNA. They are 70 nm to 90 nm in diameter and capsid is composed of 252 capsomeres. Several members of this group, including types commonly associated with respiratory disease in man, are capable of producing malignant tumors in young hamsters and a few types have been shown to be oncogenic in young rat. Previous report involving effect of caffein on transformation induced by Adenovirus type 12 [9] has been carried out. The present report represents a continuation of previous study. To obtain evidence concerning the of(tract of MNNG (N-methyl-N'-nitro-N-nitroguanidine) on transformation, investigation of adenovirus type 12 of this group was undertaken. For practical consideration it was desirable to investigate the effect of MNNG on the adenovirus type 12-induced transformation in L cell. Results were as following 1. Adeno virus type 12 induced transformation was enhanced in the presence of MNNG. 2. Yields of adeno type 12 virus in L cell were slightly inhibited by treatment of MNNG.
Adenoviridae* ; Animals ; Bedding and Linens ; Capsid ; Cricetinae ; DNA ; Methylnitronitrosoguanidine ; Rats ; Virion

Nucleocapsid protein (NPs) of negative-sense single-stranded RNA (-ssRNA) viruses function in different stages of viral replication, transcription, and maturation. Structural investigations show that -ssRNA viruses that encode NPs preliminarily serve as structural building blocks that encapsidate and protect the viral genomic RNA and mediate the interaction between genomic RNA and RNA-dependent RNA polymerase. However, recent structural results have revealed other biological functions of -ssRNA viruses that extend our understanding of the versatile roles of virally encoded NPs.
Animals ; Capsid ; metabolism ; Humans ; Lassa virus ; chemistry ; physiology ; Nucleocapsid Proteins ; chemistry ; metabolism ; Orthobunyavirus ; chemistry ; physiology ; RNA Viruses ; chemistry ; physiology

The HIV-1 capsid protein (CA) plays an essential role in viral core assembly and maturation. Proteolytic cleavage at the MA-CA junction of the retroviral gag polyprotein refolds the amino-terminal end of capsid into a beta-helix structure that is stabilized by a salt bridge between the protein's processed amino-terminus and a conserved acidic residue. The refolded capsid aminoterminus then creates a new CA-CA interface, allowing assembly of the mature capsid core. Recently, researches focus on assembly of CA in vitro and development of CA vaccine. CA vaccine will provide widely immune protection because CA is comparatively conserved. Experiments demonstrate that fusing as few as four matrix residues onto the amino-terminus of capsid redirects protein assembly from cylinder to spheres in vitro. Evaluation of immunogenicity showed that immunization with virus-like particles induced both cellular and neutralizing antibody responses. Furthermore, mucosal administration of virus-like particles effectively induced both mucosal and systemic immune responses. These results indicate that virus-like particles consisting of HIV structural proteins are an attractive vaccine platform for eliciting anti-viral immune responses, especially neutralizing antibody responses. The production of antigens for vaccines in plants indicates that plant-based transgenic expression represents a viable means of producing CA vaccine for the development of HIV vaccine and for use in HIV diagnostic procedures and it has the potential as a safe and cost-effective alternative to traditional production systems.
AIDS Vaccines ; immunology ; Capsid ; immunology ; metabolism ; Capsid Proteins ; genetics ; immunology ; metabolism ; HIV-1 ; genetics ; immunology ; metabolism ; Virion ; genetics ; immunology ; metabolism

No abstract available.
Capsid Proteins* ; Capsid*

No abstract available.
Capsid Proteins* ; Capsid* ; Genome*

For the purpose of morphologic analysis of human cytomegalovirus (HCMV) antigens reacting with specific monoclonal antibodies, we observed HCMV particles after immunogold staining. HCMV was cultured in human fetal lung fibroblasts to be concentrated by polyethylene glycol 6,000. The HCMV stock was dropped onto Formva-coated grids and was fixed by 2% glutaraldehyde. The grids were reacted with MCMVA57, 93, 135 or with SCMVM1, 6, 14, 49 monoclonal antibodies (MoAbs) follwed by gold (10 nm)-conjugated goat anti-mouse IgG. Then the grids were stained with 2.5% uranyl acetate to be observed under Hitachi 500 or Jeol 1,200 electron microscope. When HCMV was reacted with SCMVM14 and SCMVM49 MoAbs, gold particles were adsorbed to virion envelopes, suggesting that the reactive antigens were envelope proteins. In cases of MCMVA135 and SCMVM6 MoAbs, gold particles were adsorbed to dense bodies as well as to virion envelope. These results, together with the previous results of immunologic and genetic characterization, suggested that the reactive antigens of MCMVA135 and SCMVM6 MoAbs were gB homologue and structural protein, respectively. In case of SCMVM1 MoAb, gold particles were adsorbed to capsids, envelopes, and dense bodies, suggesting that the reactive antigen was structural protein. In case of SCMVM8 MoAb, gold particles were observed between the envelopes and capsids, which space was supposed to be the tegument, suggesting that the reactive antigen was carbohydrate moiety of glycoprotein or its polymer. In cases of MCMVA57 and MCMVA93 MoAbs, gold particles were adsorbed to only dense bodies, suggesting that the reactive antigens were precursors of structural proteins.
Antibodies, Monoclonal* ; Capsid ; Cytomegalovirus* ; Fibroblasts ; Glutaral ; Glycoproteins ; Goats ; Humans* ; Immunoglobulin G ; Lung ; Polyethylene Glycols ; Polymers ; Virion