BACKGROUND: Strains of respiratory syncytial virus (RSV) can be classified by reactivity with monoclonal antibodies into two major subgroups A and B, then further into several antigenic variants in each subgroup. Antigenic characterization of strains prevailing in a particular community may be essential in the future development of vaccine. METHODS: We developed monoclonal antibodies (MAbs) to RSV subgroups A and B. Antigenic specificities were characterized by immunoblot assay and radioimmunoprecipitation. By the pattern of reactions with these MAbs, RSV isolated over 1990~1998 were categorized into subgroups A and B, then further into antigenic variants. RESULTS: Thirty-four monoclonal antibodies to RSV were produced; twelve were directed against nucleoprotein; seven against matrix protein; ten against fusion protein; and five against major envelope glycoprotein. During the study period, yearly epidemics of RSV infection existed. Both subgroups circulated concurrently in 6 epidemics with predominance of subgroup A, and only one of each subgroup was identified in 2. Antigenic variations were observed in matrix, fusion and major glycoprotein. Six antigenic variants were identified in subgroup A and 2 in subgroup B among 242 strains of RSV. One major variant of subgroup A dominated in all of the 7 subgroup A epidemics, while the dominant variant of subgroup B was different in each of the 7 subgroup B epidemics. CONCLUSION: During the study period, yearly epidemics of RSV infection existed. The proportion of each subgroup varied in each of the 8 epidemics. The antigenic variability of RSV should be considered in the future development of RSV vaccine.
Antibodies, Monoclonal* ; Antigenic Variation* ; Glycoproteins ; Nucleoproteins ; Respiratory Syncytial Viruses*

Telomeres are nucleoprotein structures that compose the ends of eukaryotic chromosomes and that regulate chromosome integrity and cell proliferative lifespan. Stabilization of telomere length correlates with cell immortalization, and constitutive activation of telomerase is observed in most human cancers, suggesting that telomere maintenance plays an important role in malignant transformation and possibly aging. However, several lines of evidence indicate that alterations in telomere biology both suppress and facilitate malignant transformation. Moreover, recent observations indicate that telomerase expression plays important regulatory functions beyond the maintenance of telomere length in both normal and malignant cells. Understanding these additional functions of telomerase promise to provide further insight into both normal and malignant cell physiology.
Aging ; Biology ; Cell Physiological Phenomena ; Humans ; Nucleoproteins ; Telomerase* ; Telomere

Quantitative PCR and plaque assay are powerful virological techniques used to measure the load of defective or infectious virus in mouse and human. However, these methods display limitations such as cross contamination and long run-time. Here, we describe a novel technique termed as semi-functional quantitative flow cytometry (SFQF) for the accurate estimation of the quantity of infectious lymphocytic choriomeningitis virus (LCMV). LCMV titration method using flow cytometry was previously developed but has technical shortcomings, owing to the less optimized parameters such as cell overgrowth, plate scale, and detection threshold. Therefore, we first established optimized conditions for SFQF assay using LCMV nucleoprotein (NP)-specific antibody to evaluate the threshold of the virus detection range in the plaque assay. We subsequently demonstrated that the optimization of the method increased the sensitivity of virus detection. We revealed several new advantages of SFQF assay, which overcomes some of the previously contentious points, and established an upgraded version of the previously reported flow cytometric titration assay. This method extends the detection scale to the level of single cell, allowing extension of its application for in vivo detection of infected cells and their phenotypic analysis. Thus, SFQF assay may serve as an alternative analytical tool for ensuring the reliability of LCMV titration and can be used with other types of viruses using target-specific antibodies.
Animals ; Antibodies ; Flow Cytometry* ; Humans ; Lymphocytic choriomeningitis virus* ; Lymphocytic Choriomeningitis* ; Methods ; Mice ; Nucleoproteins ; Polymerase Chain Reaction

Rabies is a major fatal zoonotic disease in Indonesia. This study was conducted to determine the recent dynamics of rabies virus (RABV) in various areas and animal species throughout Indonesia. A total of 27 brain samples collected from rabid animals of various species in Bali, Sumatra, Kalimantan, Sulawesi, Java, and Flores in 2008 to 2010 were investigated. The cDNA of the nucleoprotein gene from each sample was generated and amplified by one-step reverse transcription-PCR, after which the products were sequenced and analyzed. The symmetric substitution model of a Bayesian stochastic search variable selection extension of the discrete phylogeographic model of the social network was applied in BEAST ver. 1.7.5 software. The spatial dispersal was visualized in Cartographica using Spatial Phylogenetic Reconstruction of Evolutionary Dynamics. We demonstrated inter-island introduction and reintroduction, and dog was found to be the only source of infection of other animals. Ancestors of Indonesian RABVs originated in Java and its descendants were transmitted to Kalimantan, then further to Sumatra, Flores, and Bali. The Flores descendent was subsequently transmitted to Sulawesi and back to Kalimantan. The viruses found in various animal species were transmitted by the dog.
Animals ; Brain ; DNA, Complementary ; Dogs ; Indonesia* ; Nucleoproteins ; Phylogeography* ; Rabies virus* ; Rabies* ; Zoonoses

China ; Genotype ; Measles virus ; genetics ; isolation & purification ; Molecular Epidemiology ; methods ; Nucleoproteins ; genetics ; Polymerase Chain Reaction

Authors studied the effects of cyciophosphamide, a potent inhibitor of nucleoprotein synthesis, to ivestigate the morphologic evidence of destructive actions to the hair. follicles. Sprague-Dawley rats were received 4 mg/kg of eyclophosphamide for 1 to 6 weeks, intraperitoneally, and examined light and electron microscopically. Light microscopically, distortion and constriction of the hair shafts, diminished diameter of the hair bulbs, and atrophy of. the hair matri.x were developed from 1 week. which were more prominent in second weeks and they were progressed after that time. Hairs were frequently fractured due to constriction of the hair shaf ts. Electron microscopically, cells of the hair pulp were decreased in number, and cells of the hair matrix were atrophied, Degenerative changes of the cellular organelles participating in grovth. and development of the hairs were noted, such as disordered formation of tricholyaline granules, diffuse atrophy and increased electron density of the inner root sheath, and loss of the glycogen and intercellular edema of the outer root sheath, but basal cells of the matrix showed minor changes relatively. From the above results, cyclophosphamide may specifically alter the matrix cells of the hair follicles and induces anagen hair losses, which may be reversible at a small dosage when the drug is discontinued, because basal cells of the matrix ahow rninor changes.
Animals ; Atrophy ; Constriction ; Cyclophosphamide* ; Edema ; Glycogen ; Hair Follicle* ; Hair* ; Nucleoproteins ; Organelles ; Rats* ; Rats, Sprague-Dawley

DNA immunization induces B and T cell responses to various pathogens and tumors. However, these responses are known to be relatively weak and often transient. Thus, novel strategies are necessary for enhancing immune responses induced by DNA immunization. Here, we demonstrated that co-immunization of influenza virus nucleoprotein (NP) gene significantly enhances humoral and cell-mediated responses to codelivered antigens in mice. We also found that NP DNA coimmunization augments in vivo proliferation of adoptively transferred antigen-specific CD4 and CD8 T cells, which enhanced protective immunity against tumor challenge. Our results suggest that NP DNA can serve as a novel genetic adjuvant in cocktail DNA vaccination.
Animals ; DNA ; Immunization ; Influenza, Human ; Mice ; Nucleoproteins ; Orthomyxoviridae ; T-Lymphocytes ; Vaccination

Rapid and reliable diagnosis of measles is important to establish an appropriate therapy and to monitor the epidemic. However, classical ELISA methods using purified virus or virus-infected cells as antigens are not only difficult to determine optimal condition for diagnosis but also highly expensive to establish routine and appropriate diagnostic systems. Nucleoprotein of measles virus is one of the major antigens of measles virus that evoke initial IgM responses. It can be used as an attractive antigen for sero-diagnosis of measles during early infection. To develop simple and inexpensive diagnostic method for measles, we expressed a recombinant nucleoprotein (60-kDa) and a fragmented portion of the nucleoprotein (50-kDa) in E. coli and evaluated their appropriateness as diagnostic antigens. The proteins strongly reacted with sera from measles patients but not with normal control sera. Efficiency of recombinant nucleoprotein-ELISA (rN-ELISA) to detect IgM was compared that of whole measles virus-ELISA (MV-ELISA) on the basis of a clinical diagnosis. In rN-ELISA, sensitivity was 73.8% and agreement was above 76.4%. In MV-ELISA, sensitivity was 76.9% and agreement was 79.2%. Therefore, efficacy of rN-ELISA seemed to be similar to that of MV-ELISA. In addition, we compared with Edmonston rN-ELISA and Korean isolate rN-ELISA on the basis of commercial MV-ELISA. In Edmonston rN-ELISA, sensitivity was 94.0% and agreement was 98.4%. In the case of Korean isolate rN-ELISA, sensitivity was 96.0% and agreement was 96.9%. Thus, there was no significant difference in the efficacy between Edmonston rN-ELISA and Korean isolate rN-ELISA. Furthermore, the correlation coefficient (R2) between Edmonston rN-ELISA and Korean isolate rN-ELISA was 0.9925. These results suggest both Edmonston and Korean isolate rN-ELISA may be useful to diagnose measles.
Diagnosis ; Enzyme-Linked Immunosorbent Assay* ; Humans ; Immunoglobulin M* ; Immunoglobulins* ; Measles virus* ; Measles* ; Nucleoproteins*

OBJECTIVETo investigate the impact of cigarette smoking on sperm nucleoprotein transition and its association with sperm motility in infertile males.

METHODSWe examined the semen quality and sperm nucleoprotein transition of 116 non-smokers and 113 heavy smokers (aged 25 -50 years) who visited the Research Institute of Obstetrics and Gynecology for male infertility. We determined the rate of individual sperm nucleoprotein transition by aniline blue staining and analyzed the correlation of cigarette smoking with routine semen parameters and the rate of sperm nucleoprotein transition. Based on the smoking index (SI) derived from smoking frequency (no. of cigarettes/d) multiplied by smoking duration (yr), the men with SI = 0 were considered as non-smokers, and those with SI > or = 200 as heavy smokers.

RESULTSThe rate of abnormal sperm nucleoprotein transition was significantly higher in the asthenozoospermic (23.5 +/- 9.4, P < 0.01) and oligoasthenozoospermic (28.2 +/- 9.2, P < 0.01) than in the normozoospermic males (19.0 +/- 9.0). Compared with the non-smokers, cigarette smoking remarkably reduced sperm nucleoprotein transition in both the men with normal sperm motility (21.9 +/- 9.8 vs 16.8 +/- 7.7, P < 0.01) and those with abnormal sperm motility (26.0 +/- 9.9 vs 22.7 +/- 8.8, P < 0.05). A weak correlation was observed between the rate of sperm nucleoprotein transition and routine semen parameters.

CONCLUSIONCigarette smoking is not significantly correlated with sperm motility but decreases sperm nucleoprotein transition in infertile males.

Adult ; Humans ; Infertility, Male ; metabolism ; Male ; Middle Aged ; Nucleoproteins ; metabolism ; Smoking ; adverse effects ; Sperm Motility ; drug effects ; Spermatozoa ; pathology

OBJECTIVETo investigate the impact of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) on sperm DNA fragmentation and nucleoprotein transition.

METHODSBased on the recommended methods in the WHO Laboratory Manual for the Examination and Processing of Human Semen (5th ed), we conducted routine semen analysis for 65 CP/CPPS patients and 30 healthy men. We also analyzed the results of papanicolaou staining, sperm DNA fragmentation and sperm nucleoprotein transition.

RESULTSCompared with the healthy control males, the CP/CPPS patients showed significant decreases in sperm concentration ([134.05 +/- 99.80] vs [94.75 +/- 92.07]) x 10(6)/ml, P <0.05), the percentage of morphologically normal sperm ([7.26 +/- 2.28] vs [5.61 +/- 3.40]%, P <0.05) and sperm progressive motility ([59.18 +/- 16.06] vs [47.68 +/- 17.62]%, P<0.05), but dramatic increases in sperm DNA fragmentation ([22.92 +/- 11.51] vs [43.58 +/- 17.07%, P<0.01) and sperm nucleoprotein transition ([23.26 +/- 5.97] vs [32.14 +/- 8.79]%, P<0.01).

CONCLUSIONCP/CPPS significantly reduces sperm quality and male fertility.

Adult ; Case-Control Studies ; DNA Fragmentation ; Humans ; Male ; Nucleoproteins ; genetics ; Prostatitis ; genetics ; Semen Analysis ; Sperm Count ; Sperm Motility ; Young Adult