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1.
Foot-and-mouth disease is an economically important viral disease in animals. It is shown that airborne diffusion is one of the main sources of contamination between animals and between herds. Epidemiological data linked to viral particle excretion can thus be used in a predictive model, added to meteorological data related to the few days before the slaughter of animals. The model computes, on a 10 km radius around the outbreak and in every space direction, the quantity of viral particles that a sensitive animal could have breathed. The aim is to define a risk area, and, according to the number and size of farms in the surrounding, to give arguments for the best sanitary decision within the emergency plan. 相似文献
2.
D Sa-Carvalho E Rieder B Baxt R Rodarte A Tanuri PW Mason 《Canadian Metallurgical Quarterly》1997,71(7):5115-5123
Isolates of foot-and-mouth disease virus (FMDV) exist as complex mixtures of variants. Two different serotype O1 Campos preparations that we examined contained two variants with distinct plaque morphologies on BHK cells: a small, clear-plaque virus that replicates in BHK and CHO cells, and a large, turbid-plaque virus that only grows in BHK cells. cDNAs encoding the capsids of these two variants were inserted into a genome-length FMDV type A12 infectious cDNA and used to produce chimeric viruses that exhibited the phenotype of the original variants. Analyses of these viruses, and hybrids created by exchanging portions of the capsid gene, identified codon 56 in VP3 (3056) as the critical determinant of both cell tropism and plaque phenotype. Specifically, the CHO growth/clear-plaque phenotype is dependent on the presence of the highly charged Arg residue at 3056, and viruses with this phenotype and genotype were selected during propagation in tissue culture. The genetically engineered Arg 3056 virus was highly attenuated in bovines, but viruses recovered from animals inoculated with high doses of this virus had lost the ability to grow in CHO cells and contained either an uncharged residue at 3056 or a negatively charged Glu substituted for a Lys at a spatially and antigenically related position on VP2 (2134). Comparison of these animal-derived viruses to other natural and engineered viruses demonstrated that positively charged residues are required at both 2134 and 3056 for binding to heparin. Taken together, these results indicate that in vitro cultivation of FMDV type O selects viruses that bind to heparin and that viruses with the heparin-binding phenotype are attenuated in the natural host. 相似文献
3.
CC Brown ME Piccone PW Mason TS McKenna MJ Grubman 《Canadian Metallurgical Quarterly》1996,70(8):5638-5641
Four calves were experimentally infected via aerosol with foot-and-mouth disease virus. Two were infected with a wild-type virus derived from a full-length infectious clone (A12-IC), and two were infected with a clone-derived virus lacking the leader gene (A12-LLV2), with euthanasia and tissue collection at 24 and 72 h postexposure (hpe). Clinical disease was apparent only in the animal given A12-IC and euthanized at 72 hpe. In situ hybridization revealed that the animal infected with A12-IC and euthanized at 24 hpe had abundant viral nucleic acid in the lung, present in clusters of positive cells in the respiratory bronchiolar epithelium and associated subepithelial regions. At 72 hpe in the A12-IC-infected calf, viral nucleic acid in the lung was present in interstitial areas, and in addition, viral nucleic acid was detectable in epithelial tissues around histologically apparent vesicles. In animals infected with A12-LLV2, viral nucleic acid was detectable in the lung at both 24 and 72 hpe, but staining revealed a more localized distribution with less nucleic acid than was found in animals given A12-IC. Therefore, it appears that after aerosol exposure to A12-IC, early replication is in the region of the lung, with subsequent dissemination to distal sites. In comparison, the A12-LLV2 virus is much less widely disseminated in the lung at 24 hpe, with no lesions or virus detectable in secondary sites at 72 hpe. The greatly reduced pathogenicity of A12-LLV2 may make it an excellent candidate for a modified live viral vaccine. 相似文献
4.
C Escarmís EC Carrillo M Ferrer JF Arriaza N Lopez C Tami N Verdaguer E Domingo MT Franze-Fernández 《Canadian Metallurgical Quarterly》1998,72(12):10171-10179
With persistent foot-and-mouth disease virus (FMDV) in BHK-21 cells, there is coevolution of the cells and the resident virus; the virulence of the virus for the parental BHK-21 cells is gradually increased, and the cells become partially resistant to FMDV. Here we report that variants of FMDV C3Arg/85 were selected in a single infection of partially resistant BHK-21 cells (termed BHK-Rb cells). Indirect immunofluorescence showed that the BHK-Rb cell population was heterogeneous with regard to susceptibility to C3Arg/85 infection. Infection of BHK-Rb cells with C3Arg/85 resulted in an early phase of partial cytopathology which was followed at 6 to 10 days postinfection by the shedding of mutant FMDVs, termed C3-Rb. The selected C3-Rb variants showed increased virulence for BHK-21 cells, were able to overcome the resistance of modified BHK-21 cells to infection, and had acquired the ability to bind heparin and to infect wild-type Chinese hamster ovary (CHO) cells. A comparison of the genomic sequences of the parental and modified viruses revealed only two amino acid differences, located at the surface of the particle, at the fivefold axis of the viral capsid (Asp-9-->Ala in VP3 and either Gly-110-->Arg or His-108-->Arg in VP1). The same phenotypic and genotypic modifications occurred in a highly reproducible manner; they were seen in a number of independent infections of BHK-Rb cells with viral preparation C3Arg/85 or with clones derived from it. Neither amino acid substitutions in other structural or nonstructural proteins nor nucleotide substitutions in regulatory regions were found. These results prove that infection of partially permissive cells can promote the rapid selection of virus variants that show alterations in cell tropism and are highly virulent for the same cells. 相似文献
5.
Growth models for predicting child stature are useful to summarize both the pattern and timing of growth in individuals and populations. Jolicoeur el al. described the JPPS model and compared it with the models of Preece and Baines (PB1) and Shohoji and Sasaki (SS). More recently Jolicoeur et al. described the JPA2 model, an extension of the JPPS. Here the JPPS model is studied in greater depth, and with more subjects, compared to the PB1 model and a modification of the SS model (SSC). The JPPS model gives consistently the best fit, although the SSC model is also better appreciably than the PB1. It is shown that biological parameters can be derived from the model parameters. Both infancy and adult data are required for the JPPS model to fit well. In some subjects the JPPS velocity curve suggests a mid-growth spurt, but it does not usually indicate a spurt in the underlying data. The SSC model is shown to be similar to Karlberg's ICP model. Overall the JPPS model provides a good fit to the child stature curve. 相似文献
6.
C Carrillo A Wigdorovitz JC Oliveros PI Zamorano AM Sadir N Gómez J Salinas JM Escribano MV Borca 《Canadian Metallurgical Quarterly》1998,72(2):1688-1690
It has been reported recently that genes encoding antigens of bacterial and viral pathogens can be expressed in plants in a form in which they retain native immunogenic properties. The structural protein VP1 of foot-and-mouth disease virus (FMDV), which has frequently been shown to contain critical epitopes, has been expressed in different vectors and shown to induce virus-neutralizing antibodies and protection in experimental and natural hosts. Here we report the production of transformed plants (Arabidopsis thaliana) expressing VP1. Mice immunized with leaf plant extracts elicited specific antibody responses to synthetic peptides representing amino acid residues 135 to 160 of VP1, to VP1 itself, and to intact FMDV particles. Additionally, all of the immunized mice were protected against challenge with virulent FMDV. To our knowledge, this is the first study showing protection against a viral disease by immunization with an antigen expressed in a transgenic plant. 相似文献
7.
Protein kinase C (PKC) is an ubiquitous regulatory enzyme with dense myocardial distribution and activity; however, its physiologic relevance to myocardial function remains poorly understood. Although endogenous Ca2+ is a potent stimulus of PKC isoforms alpha and beta (cPKCs) it remains unknown whether exogenous Ca2+ activates these PKC isoforms, and if so, whether PKC plays any role in Ca2+-induced myocardial inotropy. To study this, ventricular sections from isolated rat hearts, with and without Ca2+-induced inotropy (CaCl2, 0.5 mM coronary concentration x 2 min), were probed for cPKC isoform translocation using immunofluorescence in order to determine if exogenous Ca2+ indeed activates cPKCs. We further examined the effects of exogenous Ca2+, with and without concurrent PKC inhibition (chelerythrine, 20 microM coronary concentration x 2 min), on fundamental physiologic parameters of myocardial developed pressure (DP), dP/dt, and coronary flow (CF) in the isolated rat heart to determine if Ca2+-induced inotropy involves PKC. Results indicated that exogenous Ca2+ results in translocation of PKC a from the cytoplasm to the sarcolemma and intercalated discs, as well as the translocation of PKC beta from the perinuclear to the intranuclear compartment. This dose of exogenous Ca2+ resulted in myocardial inotropy as determined by DP, dP/dt, and CF. Furthermore, myocardial inotropy was attenuated with concurrent inhibition of PKC activity. These findings link the physiologic effects of exogenous Ca2+ to PKC, providing a better understanding of the physiologic mechanism of Ca2+-induced inotropy. 相似文献
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FJ Novembre PR Johnson MG Lewis DC Anderson S Klumpp HM McClure VM Hirsch 《Canadian Metallurgical Quarterly》1993,67(5):2466-2474
Simian immunodeficiency virus (SIV) induces an immunodeficiency syndrome similar to human AIDS. Although the disease course of SIV-induced immunodeficiency is generally measured in months to years, a disease syndrome that results in death in 5 to 14 days has been described in pig-tailed macaques infected with the SIVsmmPBj (PBj) strain. The purpose of this study was to derive an acutely lethal PBj molecular clone in order to study viral genes involved in pathogenesis. Six infectious molecular clones were generated; acutely fatal disease was induced by experimental inoculation of pig-tailed macaques with virus stocks derived from either of two clones, PBj6.6 or PBj14.6. Molecular chimeras were constructed by exchange of regions of the genome of PBj6.6 and a nonlethal, related clone, SIVsmH4. Only a chimera expressing the PBj genome under the control of a SIVsmH4 long terminal repeat induced death soon after inoculation. These studies suggest that multiple viral genes of PBj are critical for development of acute disease. More specifically, the env gene but not the long terminal repeat PBj was required for acute disease induction; however env must act in concert with another gene(s) of the PBj genome. 相似文献
11.
PG Deriabin SO Viazov EI Isaeva EI Samokhvalov DK L'vov 《Canadian Metallurgical Quarterly》1997,42(6):254-258
A model of chronic infection of primary cultures of suckling mouse brain (SMB) cells actively producing hepatitis C virus (HCV) is developed. Destruction and repopulation of cells was observed for at least 6 months; this phenomenon was paralleled by virus release in culture medium. Persistent HCV contained in SMB cultures induced a cytopathogenic effect in PS, BHK-21, Vero, HAK, and click embryo cell cultures, its infective titers being 10.0-12.0 lg TCD50/0.2 ml. Persistent HCV formed heterogeneous plaques under agar in chick embryo cells. The polymerase chain reaction (PCR) regularly detected the HCV RNA at the stage of cell destruction in the culture fluid of HCV-infected cell cultures. The cytopathogenic activity of persistent HCV was neutralized by anti-HCV positive patients' sera with the neutralization index of 8.0-9.0 lg. The results of persistent HCV neutralization were confirmed by PCR. Immunofluorescence detected virus-specific HCV antigens in 15-40% of infected cells. Hence, the SMB-HCV system realized the cytopathogenic potential of HCV circulating in the blood of patients with hepatitis C. This system is promising for the study of the pathogenesis of HCV infection at a cellular level, for screening for specific and nonspecific antiviral agents, and for preparing native virus-specific proteins and RNA. 相似文献
12.
The atmospheric dispersion of virus was simulated using a computer model which had been developed for predicting the dispersion of toxic gases from chemical engineering plants. The results were compared with data from four outbreaks in which virus was believed to have been transported by air: two outbreaks of foot-and-mouth disease in the United Kingdom in 1967 and outbreaks of Aujeszky's disease in Yorkshire in 1981 to 1982 and Indiana in 1988. There was relatively good agreement with most of these data. The paper shows that the model could be useful in an emergency because the risk of virus spread could be predicted in real time. 相似文献
13.
GR Serjeant 《Canadian Metallurgical Quarterly》1995,2(2):103-108
Amblyospora californica is a polymorphic, eukaryotic microsporidian. Three types of sporogony producing three types of spores occur in male larvae and female adults of its mosquito host, Culex tarsalis, and an alternate copepod host, Acanthocyclops vernalis. Development of A. californica in male larvae includes merogony and sporogony. Karyogamy and meiosis was observed in sporogony in male larvae but not in the female adult or in the copepod. Chromosomal evidence showed that sporogony included two consecutive meiotic divisions and a subsequent mitosis forming an octosporont, ultimately containing eight haploid, uninucleate mature spores. In this species, the haploid number of chromosomes is nine. Macrosporoblasts and macrospores, containing 1, 2 or more nuclei, can be seen in infected male larvae. The stage of sporogony in which cytokinesis was arrested seems to determine the number of nuclei. Those with only one nucleus, we believe are due to failed nuclear division at meiosis. Although A. californica displayed a process of karyogamy and meiosis similar to that of the species from Cx. salinarius, they may not be the same species because of the difference in their chromosome numbers. 相似文献
14.
Isotype-specific antibody responses to foot-and-mouth disease virus (FMDV) were measured in the sera and upper respiratory tract secretions of vaccinated and susceptible cattle challenged with FMDV by direct contact or by intranasal inoculation. A comparison was made between cattle that eliminated FMDV and those that developed and maintained a persistent infection. Serological and mucosal antibody responses were detected in all animals after challenge. IgA and IgM were detected before the development of IgG1 and IgG2 responses. IgM was not detected in vaccinated cattle. Challenge with FMDV elicited a prolonged biphasic secretory antibody response in FMDV "carrier' animals only. The response was detected as FMDV-specific IgA in both mucosal secretions and serum samples, which gained statistical significance (P < 0.05) by 5 weeks after challenge. This observation could represent the basis of a test to differentiate vaccinated and/or recovered convalescent cattle from FMDV "carriers'. 相似文献
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A major antigenic site (site A) of foot-and-mouth disease virus includes multiple overlapping epitopes located within the flexible G-H loop of capsid protein VP1. We have studied the antigenicity of several recombinant E. coli beta-galactosidases displaying the site A from a serotype C virus in different surface regions of the bacterial enzyme. In each one of the explored insertion sites, the recombinant peptide shows different specificity with a set of anti-virus monoclonal antibodies directed to site A. In some of them, the inserted stretch mimics better than free or haemocyanin-coupled peptide the antigenicity of site A in the intact virus. In particular, an insertion within an exposed loop involved in the activating interface of beta-galactosidase (amino acids 272 to 287) led to a significant improvement of the overall reactivity. Since insertions at this site renders proteins enzymatically active, the activating interface could be an adequate place for the presentation of foreign antigens in correctly assembled beta-galactosidase tetramers. These results also suggest that anti-virus antibodies directed against the major antigenic site of FMDV recognize different conformations of the G-H loop, which are better reproduced in some of the recombinant proteins because of the dissimilar restrictions imposed by each particular insertion site. 相似文献
17.
Hepatitis A virus (HAV) characteristically remains strongly cell-associated when grown in culture, with only small yields in the culture supernatant. Cell factories (6000 cm2) of BS-C-1 cells infected with the cytopathic HM175A.Z strain of HAV for 3, 4 or 7 days were harvested using trypsin to disperse the infected cell monolayer, and cells were collected by low speed centrifugation. More than 70% of the yield of virus and viral antigen can thus be obtained in the packed cell pellet. Packed cell pellets were resuspended in 5 volumes of isotonic buffer and cell membranes lysed by the addition of a non-ionic detergent. After removal of nuclei by centrifugation, ionic detergent was added to the clarified cytoplasmic extract. Under these conditions, HAV particles (virions and empty capsids) are the only particulate material remaining in the sample, and were recovered in a single ultracentrifugation step through discontinuous sucrose/glycerol density gradients. In one day, this method yields viral antigen with minimal cellular contaminants, in a concentrated volume suitable for subsequent biochemical, vaccine or diagnostic uses. The yield of viral antigen over numerous batches varied from 200 to 1600 vaccine-equivalent doses per cell factory, with a titre of up to 1 x 10(10) infectious particles per ml. 相似文献
18.
ML Marthas RA Ramos BL Lohman KK Van Rompay RE Unger CJ Miller B Banapour NC Pedersen PA Luciw 《Canadian Metallurgical Quarterly》1993,67(10):6047-6055
To identify viral determinants of simian immunodeficiency virus (SIV) virulence, two pairs of reciprocal recombinants constructed from a pathogenic (SIVmac239) and a nonpathogenic (SIVmac1A11) molecular clone of SIV were tested in rhesus macaques. A large 6.2-kb fragment containing gag, pol, env, and the regulatory genes from each of the cloned (parental) viruses was exchanged to produce one pair of recombinant viruses (designated SIVmac1A11/239gag-env/1A11 and SIVmac239/1A11gag-env/239 to indicate the genetic origins of the 5'/internal/3' regions, respectively, of the virus). A smaller 1.4-kb fragment containing the external env domain of each of the parental viruses was exchanged to create the second pair (SIVmac1A11/239env/1A11 and SIVmac239/1A11env/239) of recombinant viruses. Each of the two parental and four recombinant viruses was inoculated intravenously into four rhesus macaques, and all 24 animals were viremic by 4 weeks postinoculation (p.i.). Virus could not be isolated from peripheral blood mononuclear cells (PBMC) of any animals infected with SIVmac1A11 after 6 weeks p.i. but was consistently isolated from all macaques inoculated with SIVmac239 for 92 weeks p.i. Virus isolation was variable from animals infected with recombinant viruses; SIVmac1A11/239gag-env/1A11 and SIVmac239/1A11env/239 were isolated most frequently. Animals inoculated with SIVmac239 had 10 to 100 times more virus-infected PBMC than those infected with recombinant viruses. Three animals infected with SIVmac239 died with simian AIDS (SAIDS) during the 2-year observation period after inoculation, and the fourth SIVmac239-infected animal had clinical signs of SAIDS. Two animals infected with recombinant viruses died with SAIDS; one was infected with SIVmac239/1A11gag-env/239, and the other was infected with SIVmac1A11/239gag-env/1A11. The remaining 18 macaques remained healthy by 2 years p.i., and 13 were aviremic. One year after inoculation, peripheral lymph nodes of some of these healthy, aviremic animals harbored infected cells. All animals seroconverted within the first few weeks of infection, and the magnitude of antibody response to SIV was proportional to the levels and duration of viremia. Virus-suppressive PBMC were detected within 2 to 4 weeks p.i. in all animals but tended to decline as viremia disappeared. There was no association of levels of cell-mediated virus-suppressive activity and either virus load or disease progression. Taken together, these results indicate that differences in more than one region of the viral genome are responsible for the lack of virulence of SIVmac1A11. 相似文献
19.
The two species of the foot-and-mouth disease virus leader protein, expressed individually, exhibit the same activities 总被引:1,自引:0,他引:1
Initiation of protein synthesis on the foot-and-mouth disease virus RNA occurs at two sites, thus, two forms of the leader protein, termed Lab and Lb, are produced. Plasmids have been constructed which encode these proteins either together or individually. Plasmids encoding the Lab protein alone express a modified form of this protein in which the second methionine residue, which corresponds to the first amino acid of Lb, is changed to an alternative residue. Four different mutant forms of the Lab sequence were made. Each of the plasmids was introduced into a mammalian cell transient expression system which allowed the determination of the known activities of the L proteins. It was shown that the Lb protein and each of the modified Lab proteins were capable of cleaving the L/P1 junction in trans. Furthermore, each of these proteins induced the cleavage of the p220 component of the cap-binding complex (eIF-4F) producing inhibition of cap-dependent translation. These results indicate that the two species of L have the same functions. 相似文献
20.
Protection of swine by live and inactivated vaccines prepared from a leader proteinase-deficient serotype A12 foot-and-mouth disease virus 总被引:1,自引:0,他引:1
Previously, we demonstrated that a genetically engineered variant of foot-and-mouth disease virus (FMDV) serotype A12 lacking the leader proteinase-coding region (A12-LLV2) was attenuated and induced an immune response that partially protected cattle from FMD. In this study, A12-LLV2 was tested in swine as a live or chemically inactivated vaccine. Animals vaccinated with chemically inactivated A12-LLV2 or wild-type (WT) virus in oil adjuvant developed high levels of neutralizing antibodies and were protected from FMD upon challenge. Animals vaccinated with live A12-LLV2 did not exhibit signs of FMD, did not spread virus to other animals, developed a neutralizing antibody response and antibodies to nonstructural protein 3D, and were partially protected from FMD. Animals given a similar dose of chemically inactivated A12-LLV2 in the absence of adjuvant developed a poor immune response and were not protected from FMD, indicating that limited replication was responsible for the improved immune response found in animals vaccinated with live A12-LLV2. The results demonstrate the potential of A12-LLV2 as a live-attenuated vaccine as well as a safe source of antigen for chemically inactivated vaccines. 相似文献