[show abstract][hide abstract] ABSTRACT: Development of an electrode-modified thickness shear mode (TSM) quartz resonator that is responsive to nanogram mass loadings, while exhibiting a mass sensitivity profile that is independent of material placement on the sensor platform, is the aim of this study. The resulting nanogram balance would greatly enhance the field of mass measurement and become useful in applications such as droplet gravimetry, the study of nonvolatile residue (NVR) contamination in solvents. A ring electrode design predicted by an analytical theory for sensitivity distribution to achieve the desired uniform mass sensitivity distribution is presented in this work. Using a microvalve capable of depositing nanogram droplets of a polymer solution, and a linear stepping stage for radial positioning of these droplets across the sensor platform, measurements of the mass sensitivity distributions were conducted and are presented. The measurements agree well with theory. Further improvements are possible and are identified to achieve better uniformity and to reduce the instability in the resonant frequency of these devices. Additionally, droplet gravimetric results for NVR in methanol droplets using the modified TSM devices are presented, which compare well with determinations made by evaporation of larger volumes of the stock solutions.
[show abstract][hide abstract] ABSTRACT: This paper describes the MultiUAV2 simulation and how it has been applied to cooperative control of autonomous uninhabited air vehicles (UAV). MultiUAV2 is a simulation based on SIMULINK, Matlab, and C++ that is capable of simulating multiple UAVs which cooperate to accomplish tactical missions. First there is a discussion of cooperative control of UAVs and then the background of the MultiUAV2 simulation. Next, the simulated mission is explained, including how users can introduce new missions. Next, there are descriptions of the major elements of MultiUAV2, which are: targets/threats, vehicle dynamics, sensors, communications and cooperative assignment algorithms. In the final section, an example of the simulation event flow is presented.
American Control Conference, 2005. Proceedings of the 2005; 07/2005
[show abstract][hide abstract] ABSTRACT: Mechanistic studies of hantavirus persistence in rodent reservoirs have been limited by the lack of a versatile animal model. This report describes findings from experimental infection of inbred Lewis rats with Seoul virus strain 80-39. Rats inoculated with virus intraperitoneally at 6 days of age became persistently infected without clinical signs. Tissues from Seoul virus-inoculated 6-day-old rats were assessed at 6, 9, and 12 weeks post-inoculation for viral RNA by RT-PCR and in situ hybridization (ISH) and for infectious virus by inoculation of Vero E6 cells. Virus was isolated from lung and kidney of infected rats at 6 weeks and viral RNA was detected in lung, kidney, pancreas, salivary gland, brain, spleen, liver and skin at 6, 9 and 12 weeks. Rats inoculated with Seoul virus intraperitoneally at 10 or 21 days of age became infected without clinical signs but had low to undetectable levels of viral RNA in tissues at 6 weeks post-inoculation. ISH identified vascular smooth muscle and endothelial cells as common sites of persistent infection. Cultured rat smooth muscle cells and to a lesser extent cultured endothelial cells also were susceptible to Seoul virus infection. Pancreatic infection resulted in insulitis with associated hyperglycemia. These studies demonstrate that infant Lewis rats are uniformly susceptible to asymptomatic persistent Seoul virus infection. Additionally, they offer opportunities for correlative in vivo and in vitro study of Seoul virus interactions in host cell types that support persistent infection.
Archives of Virology 08/2004; 149(7):1325-39. · 2.03 Impact Factor
[show abstract][hide abstract] ABSTRACT: Rat coronaviruses (RCVs) are common natural pathogens of rats that cause clinical illness, necrosis, and inflammation of respiratory, salivary, and lacrimal organs. The aim of the study was to determine whether antigenically different strains of RCV vary in their pathogenic potential in rats.
Neutralization groups were identified by use of RCV strain-specific antisera. Sprague Dawley rats were inoculated oronasally with RCV-SDA, RCV-BCMM, or RCV-W. Histologic examination, immunohistochemical analysis, and reverse transcriptase-polymerase chain reaction analysis were performed on tissues from infected rats.
Clinical illness was not evident in any of the inoculated rats. The RCV-SDA strain caused mild lesions in the exorbital gland of one rat. The RCV-BCMM strain caused severe lesions in the Harderian and parotid glands and mild lesions in the exorbital glands, lungs, and nasal mucosa. The RCV-W strain caused severe lesions in the Harderian, exorbital, and parotid glands and mild lesions in the submandibular glands, lungs, and nasal mucosa. The RNA concentration was highest in the Harderian, parotid, and exorbital glands of RCV-BCMM- and RCV-W-infected rats at postinoculation day 7.
Although RCV-SDA, RCV-BCMM, and RCV-W caused different degrees and patterns of lesions, neutralization groups are not useful for predicting the pathogenic potential of a new RCV isolate.
[show abstract][hide abstract] ABSTRACT: Two serotypes of autonomously replicating parvoviruses infect laboratory mice. Genome regions coding for the nonstructural proteins of minute virus of mice [MVM] and mouse parvovirus [MPV] are almost identical, whereas capsid-coding sequences are divergent. We addressed these questions: Does humoral immunity confer protection from acute infection after challenge with homotypic or heterotypic parvovirus, and if it confers protection against acute MPV infection, does it also protect against persistent MPV infection?
Infant mice without maternal antibody or antibody to MVM or MPV and young adult mice given normal mouse serum or antibody to MVM or MPV were challenged with homotypic or heterotypic virus. In situ hybridization with target tissues was the indicator of infection.
Humoral immunity failed to confer protection against acute heterotypic parvovirus infection. In passive transfer studies, MPV DNA was observed occasionally in lymph nodes, intestine, or the spleen of MPV-challenged mice given homotypic antibody and kept for 6 or 28 days. Variable proportions of mice given MPV antibody and homotypic challenge had viral DNA in lymphoid tissues 56 days after virus inoculation.
A mouse or colony that has sustained infection with MVM or MPV is probably fully susceptible to infection with the heterotypic virus.
[show abstract][hide abstract] ABSTRACT: The recently identified autonomous mouse parvovirus designated mouse parvovirus-1 (MPV-1) persists in adult BALB/c mice for at least 9 weeks, infects lymphoid tissues, interferes with the ability of cloned T cells to proliferate, and exhibits immunomodulatory properties. As a consequence of these findings, the present studies were undertaken to characterize further the inmunomodulatory effects of MPV-1 on T cell-mediated immune responses in vivo and in vitro.
To evaluate the effect of MPV-1 infection on CD8+ T cell-mediated responses, BALB/c-H2dm2 mice were infected after transplantation of allogeneic BALB/c skin.
MPV-1 potentiated the rejection of allogeneic skin grafts. This potentiation was not a result of virus infecting the cellular or vascular component of the graft as determined by in situ hybridization, but was mediated by T cells. However, the proliferative capacity of alloantigen-reactive lymphocytes from graft-sensitized infected mice was diminished. MPV-1 also induced the rejection of syngeneic skin grafts, and T cells from these infected graft-sensitized mice lysed syngeneic P815 target cells.
These results suggest that MPV-1 infection of skin-grafted mice may disrupt normal mechanisms of peripheral tolerance and provide a unique model to study virus-induced autoimmunity.
[show abstract][hide abstract] ABSTRACT: Parvoviruses are prevalent and disruptive infectious agents of laboratory rats. Risks to rat-based research from infection are increased by the persistence of virus in immune rats and by prenatal transmission of infection. The mechanisms leading to viral persistence and prenatal infection are poorly understood and have been difficult to study for lack of reliable and humane induction methods. We report here protocols for inducing persistent and prenatal infection without causing clinical disease using the UMass strain of rat virus (RV), a common rat parvovirus. Infant rats inoculated by the oronasal route at 6 days of age had greater than 90% prevalence of persistent infection. RV-UMass also induced intrauterine infection in pregnant rats inoculated by the oronasal route. Inoculation of dams at gestation day 9 frequently caused severe disease in the fetuses whereas inoculation at gestation day 12 caused primarily asymptomatic fetal infection that persisted post partum RV-UMass infection facilitates study of parvoviralhost interactions that are relevant to laboratory rats and which also may improve understanding of persistent and prenatal human parvovirus infection.
Virus Research 10/1996; 44(1):67-78. · 2.75 Impact Factor
[show abstract][hide abstract] ABSTRACT: Parvoviruses are among the most common infectious agents of laboratory rodents and major impediments to rodent-based research. The original prototypic rodent parvoviruses-minute virus of mice, rat virus, and H-1 virus-have recently been joined by biologically and antigenically distinct parvoviruses in mice, rats, and hamsters. Recognition of the increased diversity of rodent parvoviruses presents new challenges for determining the impact of parvovirus infection on research and for detecting, preventing, and eliminating infection. This review summarizes current knowledge about rodent parvoviruses and parvovirus infections, highlighting recent research on newly isolated virus strains.
[show abstract][hide abstract] ABSTRACT: Rat coronaviruses (RCVs) infect laboratory rats and confound biomedical research results. In vitro systems developed so far have limited the growth in knowledge about RCVs by not permitting generation of plaque-cloned virus stocks, reliable isolation of RCVs from rat tissues, or growth of high titered stocks of all isolates. Due to the fact that less than 20% of L2(Percy) cells were becoming infected, sublines were produced and selected for maximal growth of RCVs. Screening of 238 cell sublines yielded L2p.176 cells which were highly susceptible to all RCVs tested; however, susceptibility declined after 30 passages in vitro. Low-passaged L2p.176 cells were used to isolate virus from natural outbreaks and to propagate individual RCV plaques into high titered stocks. Proteins from six RCV isolates were immunoblotted using polyclonal rat and mouse antibodies to sialodacryoadenitis virus and polyclonal monospecific rabbit and goat antibodies against the peplomer (S) and nucleocapsid (N) proteins of mouse hepatitis virus (MHV). Proteins of two prototype, one Japanese and three wild type RCVs were examined and found to be similar to those of MHV, although the exact sizes and ratios of protein forms were unique for most RCV isolates. This study reports the development of a continuous cell line which reliably supports RCVs opening an opportunity for further in vivo studies of the biology of these agents. As a first step in the characterization of RCVs, we have shown that RCV proteins are very similar to those of MHV.
Virus Research 04/1996; 41(1):55-68. · 2.75 Impact Factor
[show abstract][hide abstract] ABSTRACT: Lymphocytotropic mouse parvoviruses can perturb immune responses. For example the recently identified mouse parvovirus designated MPV-1 persistently infects lymphoid tissues and interferes with the ability of cloned T cells to proliferate. As a consequence of these findings the present studies were undertaken to characterize further the immunomodulatory effects of MPV-1 on T cell-mediated immune responses in vivo and in vitro. To evaluate the effect of MPV-1 on CD8+ T cell-mediated responses sarcoma I (SaI) cells, devoid of class II major histocompatibility (MHC) antigens, were administered to MPV-1-infected adult BALB/c mice. MPV-1 infection accelerated tumor allograft rejection. Immunofluorescence staining and in situ hybridization studies of tumors suggested that direct infection of the tumor cells was not responsible for accelerated rejection. Furthermore, compared with uninfected mice, T cells from infected mice that had rejected SaI tumors had a diminished cytolytic capacity. Taken together these results suggest that MPV-1 may induce "bystander help." To examine the in vivo effect of MPV-1 on CD4+ T cell mediated responses adult mice were primed with ovalbumin (OVA) and infected with MPV-1. Spleen and popliteal lymph node cells from OVA-primed mice 3 or 7 days after MPV-1 inoculation had reduced proliferation responses, whereas the proliferative capacity of mesenteric lymph node cells from these mice was increased. Similarly, MPV-1 reduced cytokine-induced proliferation of allospecific CD8+ cloned L3 T cells and OVA-reactive CD4+ T cells without effecting cell viability. Since parvoviruses are widespread among laboratory rodents, these findings emphasize the importance of identifying and excluding parvovirus infections in mice used for transplantation studies and in cultures of mouse T lymphocytes.
[show abstract][hide abstract] ABSTRACT: To characterize the serologic responses of dogs naturally exposed to or vaccinated against Borrelia burgdorferi and to assess responses at intervals after antibiotic treatment.
Prospective, controlled clinical trial.
19 dogs of various breeds and ages with narrowly defined clinical criteria of limb/joint borreliosis and 10 control dogs of equivalent age were used to determine serologic responses following natural exposure to the organism. Eight seronegative dogs were used to determine serologic responses following vaccination.
Serologic responses to B burgdorferi and recombinant outer surface protein (Osp)A, flagellin, and P39 were assessed by means of ELISA and western immunoblot. Passive protective activity was assessed by use of a mouse protection assay.
Naturally exposed dogs were seropositive, but had variable ELISA titers and immunoblot profiles. Immunoblot analysis did reveal consistent reactions to flagellin, P39, and a 22 kd protein, but not to OspA. Antibody responses did not change appreciably up to 13 weeks after antibiotic treatment. Vaccinated dogs had strong reactions to OspA and OspB, but not to P39.
Dogs with clinical borreliosis are seropositive and remain seropositive after antibiotic treatment, emphasizing that serologic testing is not a useful means of measuring clinical response. Serologic responses of infected dogs can be discriminated from those of vaccinated dogs by means of immunoblot analysis, and recombinant P39 is a potentially useful antigen for that purpose.
Journal of the American Veterinary Medical Association 01/1996; 207(11):1435-40. · 1.72 Impact Factor
[show abstract][hide abstract] ABSTRACT: An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibody to lymphocytic choriomeningitis virus (LCMV) in mouse sera. This assay is based on recombinant LCMV nucleoprotein generated in a baculovirus system. Sera from experimentally and naturally infected as well as noninfected mice were tested, and the results were compared with those obtained from an established immunofluorescence assay (IFA) that uses infected cells as antigen. An excellent correlation was found; the ELISA specificity and sensitivity were calculated to be 100 and 95% respectively. Unlike the IFA, this ELISA does not require the handling of infective virus. It eliminates the need to work with a zoonotic agent in the laboratory while allowing effective screening of laboratory mouse populations for LCMV antibody.
[show abstract][hide abstract] ABSTRACT: Inoculation of the UMass strain of rat virus (RV-UMass) into adult immunocompetent rats results in a prolonged subclinical infection that is resolved in 4 to 8 wk. Co-labeling studies, using in situ hybridization (ISH) and immunohistochemistry (IHC), confirmed that RV-UMass was lymphocytotropic and capable of infecting CD4+ and CD8+ T cells as well as B cells. ISH studies also revealed that virus replication was restricted in unstimulated cells but was productive in concanavalin A-stimulated lymphocytes. A corollary of productive infection of lymphocytes was the suppression of lymphocyte functions. Although RV-UMass did not appear to induce phenotypic changes during the course of infection, cells from infected rats had diminished proliferation and cytolytic responses. Both peripheral and mesenteric lymph node cells exhibited only partial recovery of their proliferative and cytolytic capacities one month after infection. Furthermore, RV-UMass-infected tissue culture maintained alloreactive CD4+ T cells in vitro, and a nonlethal infection of this T cell line inhibited Ag- and IL-2-induced proliferation. Because parvoviruses are widespread among laboratory rodents, these findings emphasize the importance of identifying and excluding parvovirus infection in rodents and in cultures of rat T lymphocytes.
The Journal of Immunology 11/1995; 155(8):3979-86. · 5.52 Impact Factor
[show abstract][hide abstract] ABSTRACT: In contrast to euthymic juvenile rats, which develop acute, self-limiting infection with rat virus (RV), RV infection of juvenile athymic rats was persistent for up to 12 weeks as demonstrated by recovery of infective virus, transmission to cagemates, and detection of viral DNA in the lungs. Administration of RV antiserum at the time of virus inoculation prevented persistent infection in five of six rats. Among rats given RV antiserum 1 week after virus, the interval at which euthymic rats begin to seroconvert, RV was not detected 1 week later but was recovered from four of six rats 3 weeks later. Results of these studies confirm that T-cell deficiency facilitates persistent RV infection and indicate that antibody provides significant protection from persistent infection only if it is present at the time of virus inoculation. The results support the concept that factors which prevent persistent infection in euthymic rats act early after virus inoculation and may include cellular immunity.
[show abstract][hide abstract] ABSTRACT: Infection of young adult BALB/cByJ mice with mouse parvovirus-1, a newly recognized, lymphocytotropic, nonpathogenic parvovirus, was examined by in situ hybridization. Virus appeared to enter through the small intestine and was disseminated to the liver and lymphoid tissues. Strand-specific probes detected virion DNA in a consistently larger number of cells than replicative forms of viral DNA and/or viral mRNA. The number of signal-positive cells in the intestinal mucosa, lymph nodes, spleen, and thymus increased through day 10 after oral inoculation but decreased after seroconversion. Positive cells were still detected, however, in peripheral lymphoid tissues of mice examined at 9 weeks postinoculation. The results underscore the need to assess potential effects of persistent mouse parvovirus-1 infection on immune function in mice.
Journal of Virology 07/1995; 69(6):3915-9. · 5.08 Impact Factor
[show abstract][hide abstract] ABSTRACT: C3H/He mice inoculated intradermally at one of two sites with Borrelia burgdorferi responded differently to infection. Shoulder-inoculated mice developed spirochetemia, B. burgdorferi-specific antibody, and arthritis earlier than foot-inoculated mice. Lymphocyte populations derived from spleen tissue were elevated in the shoulder- but not the foot-inoculated mice, and those from lymph nodes were increased in both groups. Lymphocytes derived from blood and spleen tissue showed impaired proliferative responses to all mitogens for shoulder-inoculated mice only, whereas proliferation of lymph node cells was not affected, regardless of route. These results demonstrate that the site of initial B. burgdorferi inoculation is an important determinant in the pathogenesis of B. burgdorferi infection.
Infection and Immunity 11/1993; 61(10):4493-7. · 4.07 Impact Factor
[show abstract][hide abstract] ABSTRACT: The oronasal median infectious doses of reovirus serotypes 1, 2, and 3 were established in infant and weanling Sencar mice on the basis of disease expression and seroconversion. Infant mice were susceptible to infection with low doses of all three serotypes, whereas weanling mice were comparatively resistant to infection. Uniform transmission of virus to cagemates or mothers of infants did not occur, indicating low contagiousness of all three virus serotypes. The comparative susceptibility of 2-day-old Sencar mice to disease was examined following oronasal inoculation with reovirus 1, 2, or 3. Tissues were collected on days 3, 5, 7, 9, 14, 16, and 21 after inoculation for virus isolation, histologic examination, and serologic analysis. Disease patterns in infant mice were distinctly different among reovirus serotypes. Reovirus 3 induced severe disease, with focal myocarditis, hepatitis, diffuse encephalitis, and generalized lymphoid depletion, whereas reovirus 1 induced a similar pattern, but much milder disease. In contrast, reovirus 2 induced mild transient enteritis without lesions in other organs. Sera from experimentally infected mice were tested in virus serotype-specific enzyme immunoassays. Cross reactivity of antibody among the three virus serotypes was found, but antibody titers were always highest with the homologous antigen. These studies confirm that infant laboratory mice are susceptible to infection with all three serotypes of virus; weanling mice are comparatively resistant to infection and disease; the viruses induce different patterns of disease in infant mice; and infecting virus serotypes can be distinguished serologically by enzyme immunoassay.
[show abstract][hide abstract] ABSTRACT: C3H/HeJ mice were inoculated intraperitoneally with 10(7) uncloned Borrelia burgdorferi at 4 weeks of age and examined on days 30, 90, 180, and 360. Spirochetes were isolated from multiple tissues at all intervals. Joint and heart disease were present in all mice at 30 days and resolved after 90 days. At 180 and 360 days, some mice had mild recurrent joint and heart disease, and most had peripheral segmental periarteritis. The protein electrophoretic migration of 360-day isolates differed from the original inoculum. The experiment was repeated with C3H/HeN and BALB/cByJ mice inoculated intradermally with 10(4) cloned B. burgdorferi. Characterization of infection and disease at 180 and 360 days were similar to those of the first experiment, but spirochetal proteins of isolates from both intervals displayed no protein variation in electrophoretic mobilities. Spirochetes isolated at 360 days were fully pathogenic in naive mice. Sera from infected mice showed an initial immunoglobulin M response, followed by a sustained immunoglobulin G response, involving IgG1, IgG2a, IgG2b and IgG3, with expanding reactivity against multiple antigens over time. These results indicate that immunocompetent mice sustain persistent infections and develop early acute joint and heart lesions that resolve and then recur intermittently.
American Journal Of Pathology 10/1993; 143(3):959-71. · 4.52 Impact Factor
[show abstract][hide abstract] ABSTRACT: The course of enterotropic mouse hepatitis virus (MHV) infection was examined in genetically susceptible (BALB) and resistant (SJL) mice of different ages at inoculation (1, 3, and 12 weeks) and at sequential intervals (1, 2, 3, 5, 10, 20, and 30 days) after oral inoculation with the Y strain of MHV (MHV-Y). Virus was quantified in stomach, upper and lower segments of small intestine, cecum, upper and lower segments of colon, Peyer's patches, mesenteric lymph node, and feces, and tissues were examined microscopically. An infant mouse bioassay was used to quantify virus in all tissues of 3-week-old BALB mice and ascending colons of other mouse groups. MHV-specific serum IgG antibody titers were measured with an enzyme immunoassay, using MHV-S-infected 17 Cl 1 cells as antigen. Lesions were first detectable at 2 days and were most severe in 1-week-old mice and more severe in BALB mice, compared with SJL mice of the same age. Additional BALB mice inoculated at the age of 24 hours developed severe necrotizing enterocolitis, whereas SJL mice inoculated at the age of 24 hours developed lesions equivalent to those in 1-week-old BALB mice. Virus was first detectable at 2 days and virus titers were highest at 2, 3, and 5 days, then diminished on days 10, 20, and 30. Low titers of virus were found in a few mice of different ages and genotypes through day 30. Lesions were most severe and virus titers highest in the ascending colon.(ABSTRACT TRUNCATED AT 250 WORDS)