bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neurons Differentially Activate the Herpes Simplex Virus Type 1 Immediate-Early Gene ICP0 and ICP27 Promoters in Transgenic Mice
Authors: Loiacono Christie M. , Myers Robert , Mitchell William J. 
Published: July 2002

ABSTRACT

Herpes simplex virus type 1 (HSV-1) immediate-early (IE) proteins are required for the expression of viral early and late proteins. It has been hypothesized that host neuronal proteins regulate expression of HSV-1 IE genes that in turn control viral latency and reactivation. We investigated the ability of neuronal proteins in vivo to activate HSV-1 IE gene promoters (ICP0 and ICP27) and a late gene promoter (gC). Transgenic mice containing IE (ICP0 and ICP27) and late (gC) gene promoters of HSV-1 fused to the
Escherichia coli
β-galactosidase coding sequence were generated. Expression of the ICP0 and ICP27 reporter transgenes was present in anatomically distinct subsets of neurons in the absence of viral proteins. The anatomic locations of β-galactosidase-positive neurons in the brains of ICP0 and ICP27 reporter transgenic mice were similar and included cerebral cortex, lateral septal nucleus, cingulum, hippocampus, thalamus, amygdala, and vestibular nucleus. Trigeminal ganglion neurons were positive for β-galactosidase in adult ICP0 and ICP27 reporter transgenic mice. The ICP0 reporter transgene was differentially regulated in trigeminal ganglion neurons depending upon age. β-Galactosidase-labeled cells in trigeminal ganglia and cerebral cortex of ICP0 and ICP27 reporter transgenic mice were confirmed as neurons by double labeling with antineurofilament antibody. Nearly all nonneuronal cells in ICP0 and ICP27 reporter transgenic mice and all neuronal and nonneuronal cells in gC reporter transgenic mice were negative for β-galactosidase labeling in the absence of HSV-1. We conclude that factors in neurons are able to differentially regulate the HSV-1 IE gene promoters (ICP0 and ICP27) in transgenic mice in the absence of viral proteins. These findings are important for understanding the regulation of the latent and reactivated stages of HSV-1 infection in neurons.
Budding of Equine Infectious Anemia Virus Is Insensitive to Proteasome Inhibitors
Authors: Patnaik Akash , Chau Vincent , Li Feng , Montelaro Ronald C. , Wills John W. 
Published: July 2002

ABSTRACT
The only retrovirus protein required for the budding of virus-like particles is the Gag protein; however, recent studies of Rous sarcoma virus (RSV) and human immunodeficiency virus have suggested that modification of Gag with ubiquitin (Ub) is also required. As a consequence, the release of these viruses is reduced in the presence of proteasome inhibitors, which indirectly reduce the levels of free Ub within the cell. Here we show that the budding of equine infectious anemia virus (EIAV) from infected equine cells is largely unaffected by these drugs, although use of one inhibitor (MG-132) resulted in a dramatic block to proteolytic processing of Gag. This lack of sensitivity was also observed in transiently transfected avian cells under conditions that greatly reduce RSV budding. Moreover, insensitivity was observed when the EIAV Gag protein was expressed in the absence of all the other virus products, indicating that they are not required for this phenotype. An activity that enables EIAV to tolerate exposure to proteasome inhibitors was mapped to the C-terminal p9 sequence, as demonstrated by the ability of an RSV Gag-p9 chimera to bud in the presence of the drugs. Intriguingly, the p9 sequence contains a short sequence motif that is similar to a surface-exposed helix of Ub, suggesting that EIAV Gag may have captured a function that allows it to bypass the need for ubiquitination. Thus, the mechanism of EIAV budding may not be substantially different from that of other retroviruses, even though it behaves differently in the presence of proteasome inhibitors.
Novel, Chimpanzee Serotype 68-Based Adenoviral Vaccine Carrier for Induction of Antibodies to a Transgene Product
Authors: Xiang Zhiquan , Gao Guangping , Reyes-Sandoval Arturo , Cohen Christopher J. , Li Yan , Bergelson Jeffrey M. , Wilson James M. , Ertl Hildegund C. J. 
Published: July 2002

ABSTRACTAn E1-deletion-containing adenoviral recombinant based on the chimpanzee serotype 68 (AdC68) was developed to express the rabies virus glycoprotein. Mice immunized with this construct (AdC68rab.gp) developed antibodies to rabies virus and remained resistant to challenge with an otherwise lethal dose of rabies virus. In naïve mice immunized intranasally, the rabies virus-specific antibody responses elicited by AdC68rab.gp were comparable with regard to both titers and isotype profiles to those induced by an adenoviral recombinant based on human serotype 5 (Adhu5) expressing the same transgene product. In contrast, subcutaneous immunization with the AdC68rab.gp vaccine resulted in markedly lower antibody responses to the rabies virus glycoprotein than the corresponding Adhu5 vaccine. Antibodies from AdC68rab.gp-immunized mice were strongly biased towards the immunoglobulin G2a isotype. The antibody response to the rabies virus glycoprotein presented by Adhu5rab.gp was severely compromised in animals preexposed to the homologous adenovirus. In contrast, the rabies virus-specific antibody response to the AdC68rab.gp vaccine was at most marginally affected by preexisting immunity to common human adenovirus serotypes, such as 2, 4, 5, 7, and 12. This novel vaccine carrier thus offers a distinct advantage over adenoviral vaccines based on common human serotypes.
Improving DNA Vaccine Potency by Linking Marek's Disease Virus Type 1 VP22 to an Antigen
Authors: Hung Chien-Fu , He Liangmei , Juang Jeremy , Lin Tzyy-Jye , Ling Morris , Wu T.-C. 
Published: July 2002

ABSTRACTWe have previously employed an intercellular spreading strategy using herpes simplex virus type 1 (HSV-1) VP22 protein to enhance DNA vaccine potency because DNA vaccines lack the intrinsic ability to amplify in cells. Recently, studies have demonstrated that the protein encoded by UL49 of Marek's disease virus type 1 (MDV-1) exhibits some degree of homology to the HSV-1 VP22 protein and features the property of intercellular transport. We therefore generated a DNA vaccine encoding MDV-1 VP22 linked to a model antigen, human papillomavirus type 16 E7. We demonstrated that compared with mice vaccinated with DNA encoding wild-type E7, mice vaccinated with MDV-1 VP22/E7 DNA exhibited a significant increase in number of gamma-interferon-secreting, E7-specific CD8+-T-cell precursors as well as stronger tumor prevention and treatment effects. Furthermore, our data indicated that the antitumor effect was CD8 dependent. These results suggested that the development of vaccines encoding VP22 fused to a target antigen might be a promising strategy for improving DNA vaccine potency.
Novel Swine Virulence Determinant in the Left Variable Region of the African Swine Fever Virus Genome
Authors: Neilan J. G. , Zsak L. , Lu Z. , Kutish G. F. , Afonso C. L. , Rock D. L. 
Published: July 2002

ABSTRACT

Previously we have shown that the African swine fever virus (ASFV)
NL
gene deletion mutant E70ΔNL is attenuated in pigs. Our recent observations that
NL
gene deletion mutants of two additional pathogenic ASFV isolates, Malawi Lil-20/1 and Pr4, remained highly virulent in swine (100% mortality) suggested that these isolates encoded an additional virulence determinant(s) that was absent from E70. To map this putative virulence determinant, in vivo marker rescue experiments were performed by inoculating swine with infection-transfection lysates containing E70
NL
deletion mutant virus (E70ΔNL) and cosmid DNA clones from the Malawi
NL
gene deletion mutant (MalΔNL). A cosmid clone representing the left-hand 38-kb region (map units 0.05 to 0.26) of the MalΔNL genome was capable of restoring full virulence to E70ΔNL. Southern blot analysis of recovered virulent viruses confirmed that they were recombinant E70ΔNL genomes containing a 23- to 28-kb DNA fragment of the Malawi genome. These recombinants exhibited an unaltered MalΔNL disease and virulence phenotype when inoculated into swine. Additional in vivo marker rescue experiments identified a 20-kb fragment, encoding members of multigene families (MGF) 360 and 530, as being capable of fully restoring virulence to E70ΔNL. Comparative nucleotide sequence analysis of the left variable region of the E70ΔNL and Malawi Lil-20/1 genomes identified an 8-kb deletion in the E70ΔNL isolate which resulted in the deletion and/or truncation of three MGF 360 genes and four MGF 530 genes. A recombinant MalΔNL deletion mutant lacking three members of each MGF gene family was constructed and evaluated for virulence in swine. The mutant virus replicated normally in macrophage cell culture but was avirulent in swine. Together, these results indicate that a region within the left variable region of the ASFV genome containing the MGF 360 and 530 genes represents a previously unrecognized virulence determinant for domestic swine.
Sustained High Frequencies of Specific CD4 T Cells Restricted to a Single Persistent Virus
Authors: Sester Martina , Sester Urban , Gärtner Barbara , Kubuschok Boris , Girndt Matthias , Meyerhans Andreas , Köhler Hans 
Published: July 2002

ABSTRACTReplication of cytomegalovirus (CMV) is largely controlled by the cellular arm of the immune response. In this study the CMV-specific CD4 T-cell response was characterized in a cohort of apparently healthy individuals. In 11% of all individuals, extremely high frequencies, between 10 and 40%, were found. High-level frequencies of CMV-specific CD4 T cells persisted over several months and were not the result of an acute infection. Specific T cells were oligoclonal and were phenotypically and functionally characterized as mature effector cells, with both cytokine-secreting and proliferative potential. These high-level frequencies do not seem to compromise the immune response towards heterologous infections, and no signs of immunopathology were observed. Whereas a large temporary expansion of virus-specific T cells is well known to occur during acute infection, we now show that extremely high frequencies of virus-specific T cells may continuously exist in chronic CMV infection without overtly compromising the remaining protective immunity.
Reovirus Core Protein μ2 Determines the Filamentous Morphology of Viral Inclusion Bodies by Interacting with and Stabilizing Microtubules
Authors: Parker John S. L. , Broering Teresa J. , Kim Jonghwa , Higgins Darren E. , Nibert Max L. 
Published: July 2002

ABSTRACTCells infected with mammalian reoviruses often contain large perinuclear inclusion bodies, or “factories,” where viral replication and assembly are thought to occur. Here, we report a viral strain difference in the morphology of these inclusions: filamentous inclusions formed in cells infected with reovirus type 1 Lang (T1L), whereas globular inclusions formed in cells infected with our laboratory's isolate of reovirus type 3 Dearing (T3D). Examination by immunofluorescence microscopy revealed the filamentous inclusions to be colinear with microtubules (MTs). The filamentous distribution was dependent on an intact MT network, as depolymerization of MTs early after infection caused globular inclusions to form. The inclusion phenotypes of T1L × T3D reassortant viruses identified the viral M1 genome segment as the primary genetic determinant of the strain difference in inclusion morphology. Filamentous inclusions were seen with 21 of 22 other reovirus strains, including an isolate of T3D obtained from another laboratory. When the μ2 proteins derived from T1L and the other laboratory's T3D isolate were expressed after transfection of their cloned M1 genes, they associated with filamentous structures that colocalized with MTs, whereas the μ2 protein derived from our laboratory's T3D isolate did not. MTs were stabilized in cells infected with the viruses that induced filamentous inclusions and after transfection with the M1 genes derived from those viruses. Evidence for MT stabilization included bundling and hyperacetylation of α-tubulin, changes characteristically seen when MT-associated proteins (MAPs) are overexpressed. Sequencing of the M1 segments from the different T1L and T3D isolates revealed that a single-amino-acid difference at position 208 correlated with the inclusion morphology. Two mutant forms of μ2 with the changes Pro-208 to Ser in a background of T1L μ2 and Ser-208 to Pro in a background of T3D μ2 had MT association phenotypes opposite to those of the respective wild-type proteins. We conclude that the μ2 protein of most reovirus strains is a viral MAP and that it plays a key role in the formation and structural organization of reovirus inclusion bodies.
Photophysics of Cyanine Dyes on Surfaces: Laser-Induced Photoisomer Emission of 3,3'-Dialkylthiacarbocyanines Adsorbed on Microcrystalline Cellulose
Authors: Oliveira Anabela S. , Almeida Paulo , Vieira Ferreira Luis Filipe 
Published: July 2002

The photophysics of three thiacarbocyanine dyes, 3,3'-dimethylthiacarbocyanine iodide (DMTCC), 3,3'-diethylthiacarbocyanine iodide (DETCC), and 3,3′-dipropylthiacarbocyanine iodide (DPTCC) was studied when adsorbed on microcrystalline cellulose in the concentration range from 5.0 · 10-4 to 10.0 μmol g-1. Using ground-state diffuse reflectance absorption technique, only H aggregate formation was detected for all the probes. The amount of aggregate formed depends on the hydration degree of the sample, always decreasing with sample dryness. The fluorescence quantum yields for all the adsorbed dyes are one order of magnitude higher than those observed in nonviscous solvents, being 0.98 for DMTCC, 0.96 for DETCC, and 0.63 for DPTCC. Laser-induced fluorescence emissions were recorded (using an intensified-charge-coupled-device detection system) as a function of the laser power, showing that for dry concentrated samples irradiated with high laser intensity, a second fluorescence emission band (bathochromically shifted relative to the monomer emission) was detected. This emission shows a supralinear dependence on laser power. The new emissions here detected arise from fluorescent photoisomers formed via singlet monomers, by a two-photon absorption process.
Should infants be screened for anaemia? A prospective study investigating the relation between haemoglobin at 8, 12, and 18 months and development at 18 months
Authors: Sherriff A , Emond A , Bell J C , Golding J , Team the ALSPAC Study 
Published: July 2002

AIMSTo investigate the relation between haemoglobin in children followed longitudinally from 8 to 18 months, and developmental outcome at 18 months.METHODSThe Avon Longitudinal Study of Pregnancy and Childhood (ALSPAC) is a longitudinal survey of a geographically defined population of children born in 1991–92. In a randomly selected subsample, blood samples were assayed for Hb at 8, 12, and 18 months; a developmental assessment was carried out at 18 months on 1141 children using the Griffiths Scales of Mental Development.RESULTSThere was a strong quadratic association between Hb at 8 months and performance on the locomotor subscale at 18 months. Average scores increased with increasing Hb up to 95 g/l; there was little additional developmental benefit in Hb levels beyond 95 g/l. Infants with Hb <95 g/l at 8 months of age scored on average 6 points lower on the locomotor subscale than infants with Hb ⩾ 95 g/l; infants with Hb <90 g/l at 8 months scored 12 points lower on the locomotor subscale than children with Hb ⩾ 90 g/l.CONCLUSIONSLow Hb concentrations (⩽ 95 g/l) in 8 month old children are associated with impaired motor development at 18 months. This cut off point corresponds to the 5th centile of Hb at 8 months. The results indicate that if there is an adverse effect of low Hb on developmental outcome, screening may be more effective at 8 months or earlier, rather than after this age. We propose to examine the importance of infant anaemia in relation to more accurate and detailed long term outcomes as the children get older.
Physiological and metabolic characteristics of elite tug of war athletes
Authors: Warrington G , Ryan C , Murray F , Duffy P , Kirwan J P 
Published: July 2002

Objective—To determine the aerobic power (V̇o2max), body composition, strength, muscular power, flexibility, and biochemical profile of an elite international squad of tug of war athletes.Methods—Sixteen male competitors (mean (SEM) age 34 (2) years) were evaluated in a laboratory. For comparative purposes, data were analysed relative to normative data for our centre and to a group of 20 rugby forwards from the Irish international squad.Results—The tug of war participants were lighter (83.6 (3.0)v104.4 (1.8) kg, p<0.0001) and had less lean body mass (69.4 (2.1)v86.2 (1.2) kg) than the rugby players and had lower than normal body fat (16.7 (0.9)%); all values are mean (SEM). Aerobic power measured during a treadmill test was 55.8 (1.6) ml/kg/min for the tug of war participants compared with 51.1 (1.4) ml/kg/min for the rugby forwards (p<0.03). A composite measure of strength derived from (sum of dominant and non-dominant grip strength and back strength)/lean body mass yielded a strength/mass ratio that was 32% greater (p<0.0001) for the tug of war group than the rugby group. Dynamic leg power was lower for the tug of war group than the rugby forwards (4659.8 (151.6)v6198.2 (105) W respectively; p<0.0001). Leg flexibility was 25.4 (2.0) cm for the tug of war group. Back flexibility was 28.6 (1.4) cm which was lower (p<0.02) than the rugby forwards 34.2 (1.5) cm. Whereas blood chemistry and haematology were normal, packed cell volume, haemoglobin concentration, and erythrocyte volume were lower in the tug of war group than in the rugby players (p<0.05). All three haematological measures correlated with muscle mass (packed cell volume,r2= 0.37, p<0.0001; haemoglobin concentration,r2= 0.13, p<0.05; erythrocyte volume,r2= 0.21, p<0.01).Conclusions—The data indicate that international level tug of war participants have excellent strength and above average endurance relative to body size, but have relatively low explosive leg power and back flexibility. The data provide reference standards for the sport and may be useful for monitoring and evaluating current and future participants.
Teaching old dogs new tricks—a personal perspective on a decade of efforts by a clinical ethics committee to promote awareness of medical ethics
Authors: Tweeddale Martin G 
Published: July 2002

To incorporate medical ethics into clinical practice, it must first be understood and valued by health care professionals. The recognition of this principle led to an expanding and continuing educational effort by the ethics committee of the Vancouver General Hospital. This paper reviews this venture, including some pitfalls and failures, as well as successes. Although we began with consultants, it quickly became apparent that education in medical ethics must reach all health care professionals—and medical students as well. Our greatest successes came in the formative years of a medical career (ie, in medical school and residency training programmes), but other efforts were not wasted, particularly among nurses and other health care professionals.Although this is a personal review of the experience in one institution, the lessons learnt in Vancouver are applicable to the further development of medical ethics in the UK.
Moderate sleep deprivation produces impairments in cognitive and motor performance equivalent to legally prescribed levels of alcohol intoxication
Authors: Williamson A M , Feyer Anne-Marie 
Published: July 2002

OBJECTIVESTo compare the relative effects on performance of sleep deprivation and alcohol.METHODSPerformance effects were studied in the same subjects over a period of 28 hours of sleep deprivation and after measured doses of alcohol up to about 0.1% blood alcohol concentration (BAC). There were 39 subjects, 30 employees from the transport industry and nine from the army.RESULTSAfter 17–19 hours without sleep, corresponding to 2230 and 0100, performance on some tests was equivalent or worse than that at a BAC of 0.05%. Response speeds were up to 50% slower for some tests and accuracy measures were significantly poorer than at this level of alcohol. After longer periods without sleep, performance reached levels equivalent to the maximum alcohol dose given to subjects (BAC of 0.1%).CONCLUSIONSThese findings reinforce the evidence that the fatigue of sleep deprivation is an important factor likely to compromise performance of speed and accuracy of the kind needed for safety on the road and in other industrial settings.
Living near opencast coal mining sites and children's respiratory health
Authors: Pless-Mulloli Tanja , Howel Denise , King Andrew , Stone Ian , Merefield John , Bessell Jan , Darnell Ross 
Published: July 2002

OBJECTIVESTo answer the question whether living near opencast coal mining sites affects acute and chronic respiratory health.METHODSAll 4860 children aged 1–11 from five socioeconomically matched pairs of communities close to active opencast sites and control sites away from them were selected. Exposure was assessed by concentrations of particulate matter with aerodynamic diameter <10 μm (PM10), residential proximity to active opencast sites, and particle composition. PM10was monitored and sampled for 6 weeks in four pairs, and for 24 weeks in one pair. A postal questionnaire collected data on health and lifestyle. Daily health information was collected by a symptom diary (concurrently with PM10monitoring) and general practitioner (GP) records were abstracted (concurrently with PM10monitoring and 52 weeks before the study). Outcomes were the cumulative and period prevalence (2 and 12 months) of wheeze, asthma, bronchitis, and other respiratory symptoms, and the prevalence and incidence of daily symptoms and GP consultations.RESULTSPatterns of the daily variation of PM10were similar in opencast and control communities, but PM10was higher in opencast areas (mean ratio 1.14, 95% confidence interval (95% CI) 1.13 to 1.16, geometric mean 17.0 μg/m3v14.9 μg/m3). Opencast sites were a measurable contributor to PM10in adjacent areas. Little evidence was found for associations between living near an opencast site and an increased prevalence of respiratory illnesses, asthma severity, or daily diary symptoms, but children in opencast communities 1–4 had significantly more respiratory consultations (1.5v1.1 per person-year) than children in control communities for the 6 week study periods. Associations between daily PM10concentrations and acute health events were similar in opencast and control communities.CONCLUSIONSChildren in opencast communities were exposed to a small but significant amount of additional PM10to which the opencast sites were a measurable contributor. Past and present respiratory health of children was similar, but GP consultations for respiratory conditions were higher in opencast communities during the core study period.