bluuebunny/crossref_metadata_2025 huggingface dataset:
Low‐field standing magnetic resonance imaging findings of the metacarpo/metatarsophalangeal joint of racing Thoroughbreds with lameness localised to the region: A retrospective study of 131 horses
Authors: POWELL S. E.
Published: June 2011

Reasons for performing study:The metacarpo/metatarsophalangeal (MCP/MTP) joint is a common site of lameness in the Thoroughbred racehorse. Radiographs may fail to show pathology consistent with the degree of lameness. With a high incidence of stress fractures occurring in the distal third metacarpal/metatarsal (MC3/MT3) condyles and proximal phalanx, a definitive diagnosis as to the nature of the pathology is essential.

Objective:To describe the low‐field magnetic resonance imaging (MRI) findings in Thoroughbred racehorses with MCP/MTP joint pain scanned under standing sedation.

Methods:The MR images and clinical records of all Thoroughbred racehorses undergoing MRI of the MCP/MTP joint between October 2006 and August 2010 were reviewed. A total of 168 joints from 131 horses were selected for inclusion. The MRI finding considered most significant in the lame (or lamest) limb was noted.

Results:Diagnostic quality images were obtainable in 97.8% of horses. The most common finding was palmar/plantar osteochondral disease in MC3/MT3 (54.9% of horses). Incomplete condylar fracture was diagnosed in 19.8% of horses, with the lateral condyle predominating. An incomplete sagittal fracure of P1 was diagnosed in 14.5% of horses and 11.4% were diagnosed with ‘dorsal joint disease’. Other findings included transverse MC3 stress fractures (1.5%), soft tissue injuries (12.2%) and proximal phalangeal ‘contusions’ (3.8%). No significant bone/soft tissue injury was detected in 5.4% of cases.

Conclusions:Standing MRI can detect a spectrum of disease within the MCP/MTP joints of racehorses. The procedure is well tolerated and may lead to a definitive diagnosis where radiographic imaging is inconclusive.

Potential relevance:A total of 35.8% of cases had MRI findings consistent with fracture pathology, which could not be confirmed radiographically at the time of the MRI examination. This has important therapeutic and prognostic implications and may help to prevent catastrophic injury.
Phacoemulsification and implantation of foldable +14 diopter intraocular lenses in five mature horses
Authors: TOWNSEND W. M. , JACOBI S. , BARTOE J. T.
Published: June 2011

Presently, intraocular lenses (IOLs) are not routinely implanted after equine cataract surgery. Subsequently, horses are visual but markedly farsighted (hyperopic). This report describes the surgical results and visual status after phacoemulsification and implantation of IOLs in mature horses with spontaneous cataracts. Six eyes of 5 mature horses underwent phacoemulsification and implantation of a +14 diopter (D) foldable IOL. Recheck ocular examinations were performed at 1, 4 and 24 weeks post operatively. Refractive error was recorded at 4 weeks post operatively. Visual status, refractive error and anterior chamber depth were recorded 24 weeks post operatively: 5 of 6 operated eyes remained visual and the average refractive error was +0.4 ± 1.1D. There was a significant difference between the 24 week post operative refractive error and the population mean of +10D (P<0.0001) for aphakic horses. The average post operative anterior chamber depth was 7.89 ± 1.55 mm. One globe was enucleated 2 months post operatively. Phacoemulsification with IOL implantation resulted in a significant reduction in post operative refractive error and restored vision to within 0.4D of emmetropia in 5 of 6 operated eyes. Implantation of a +14D IOL ameliorated the hyperopia documented in aphakic horses and improved the post operative visual acuity.
Comparison of owner‐reported health problems with veterinary assessment of geriatric horses in the United Kingdom
Authors: IRELAND J. L. , CLEGG P. D. , McGOWAN C. M. , McKANE S. A. , CHANDLER K. J. , PINCHBECK G. L.
Published: June 2011

Reasons for performing study: Previous studies suggest that owners underestimate or incorrectly recognise or report health problems in geriatric horses. However, few studies have directly compared owner‐reported and veterinary assessed disease.

Objectives: To compare the findings of veterinary clinical examination of geriatric horses with owner‐reported clinical signs and disease.

Methods: A total of 200 horses aged ≥15 years were randomly selected to receive a veterinary examination, from responses to a cross‐sectional postal questionnaire survey. Veterinary examinations were performed within 2 months of questionnaire return, and agreement between owner‐reported data and veterinary clinical findings was assessed.

Results: Owners under‐reported many clinical signs and disease conditions detected on veterinary clinical examination. For example, dental abnormalities (detected in 95.4% of horses, reported by 24.5% of owners); cardiac murmurs (detected in 20% of horses, reported by 0.5% of owners); lameness (present in 50% of horses, reported by 23% of owners) and hoof abnormalities (detected in 80% of horses, reported by 27% of owners). Agreement between owner‐reported and veterinary assessed respiratory disease (Kappa 0.02–0.2), body condition score (Kappa 0.24) and coat abnormalities (Kappa 0.42) was poor, fair and moderate, respectively. Range of motion (ROM) of the tarsal and metacarpophalangeal joints was lower in horses with owner‐reported osteoarthritis (P = 0.005 and <0.001, respectively).

Conclusions and potential relevance: The low prevalence and relatively poor agreement of owner‐reported disease compared to that detected on veterinary examination suggests inaccurate or under‐recognition, or inaccurate reporting of health problems by owners of geriatric horses, which could lead to a delay in presentation for veterinary treatment. Increased veterinary involvement and improved owner education in the care of geriatric horses should facilitate earlier identification of disease, particularly that which is not readily detectable by owners, and aid management of health and welfare problems.
Cervical vertebral canal endoscopy in a horse with cervical vertebral stenotic myelopathy
Authors: PRANGE T. , CARR E. A. , STICK J. A. , GARCIA‐PEREIRA F. L. , PATTERSON J. S. , DERKSEN F. J.
Published: June 2011

A 3‐year‐old Thoroughbred gelding presented with a history of neurological signs, including incoordination in his hindlimbs, of about 7 months' duration. On initial examination, the horse exhibited ataxia and paresis in all limbs with more severe deficits in the hindlimbs. Cervical radiographs displayed severe osteoarthritis of the articular processes between C5 and C6. On subsequent cervical myelography the dorsal contrast column was reduced by 90% at the level of the intervertebral space between C5 and C6. Cervical vertebral canal endoscopy, including epidural (epiduroscopy) and subarachnoid endoscopy (myeloscopy), was performed under general anaesthesia. A substantial narrowing of the subarachnoid space at the level between C6 and C7 was seen during myeloscopy, while no compression was apparent between C5 and C6. Epiduroscopy showed no abnormalities. After completion of the procedure, the horse was subjected to euthanasia and the cervical spinal cord submitted for histopathological examination. Severe myelin and axon degeneration of the white matter was diagnosed at the level of the intervertebral space between C6 and C7, with Wallerian degeneration cranially and caudally, indicating chronic spinal cord compression at this site. Myeloscopy was successfully used to identify the site of spinal cord compression in a horse with cervical vertebral stenotic myelopathy, while myelography results were misleading.
Short‐term temporal alterations in magnetic resonance signal occur in primary lesions identified in the deep digital flexor tendon of the equine digit
Authors: MILNER P. I. , SIDWELL S. , TALBOT A. M. , CLEGG P. D.
Published: June 2011

Reasons for performing study: Primary lesions of the deep digital flexor tendon (DDFT) within the digit are an important cause of lameness diagnosed using magnetic resonance imaging (MRI) but appearance of these lesions over time has not been documented.

Objectives: To determine whether the magnetic resonance (MR) appearance of different primary DDFT lesions alter over a 6 month period and whether lesion type is a determinant of these changes.

Methods: Cases included had lameness attributable to a primary lesion involving the DDFT in the digit diagnosed on MRI. Lesions were typed into parasagittal, dorsal border and core lesions. Approximate volumes and intensities were quantified for each lesion type using T2* scan sequences. Follow‐up examinations and measurements were repeated at 3 and 6 month periods following conservative management.

Results: Twenty‐three horses fitted the inclusion criteria. Lesion distribution included: parasagittal (n = 7), dorsal border (n = 11) and core lesions (n = 5). No association was found between age of horse, degree of lameness and lesion type. Only dorsal border lesions showed statistically significant reduction both in volume (initial scan: 0.18 ± 0.14 cm3) at 3 months (0.11 ± 0.10 cm3, P<0.05) and 6 months (0.05 ± 0.05 cm3, P<0.01) and ratiometric intensity (initial scan: 4.06 ± 1.54) at 6 months (2.00 ± 0.43; P<0.01). Parasagittal and core lesions showed no difference in lesion volume or ratiometric intensity. Lameness improved in all lesion types following conservative management.

Conclusions: Dorsal border lesions of the DDFT show reduction in both volume and intensity whereas parasagittal and core lesions do not.

Potential relevance: Lesion typing may be important in predicting lesion behaviour and short‐term outcome using MR imaging.
Serum concentration of surfactant protein D in horses with lower airway inflammation
Authors: RICHARD E. A. , PITEL P.‐H. , CHRISTMANN U. , LEKEUX P. , FORTIER G. , PRONOST S.
Published: June 2011

Reasons for performing study:Surfactant protein D (SP‐D), mainly synthesised by alveolartype IIcells and nonciliated bronchiolar cells, is one important component of innate pulmonary immunity. In man, circulating concentrations of SP‐D are routinely used as biomarkers for pulmonary injury. To date, serum SP‐D levels have only been investigated in horses in an experimental model of bacterial airway infection.

Objectives:To compare serum SP‐D concentrations at rest and after exercise in horses with and without inflammatory airway disease (IAD).

Methods:Venous blood samples were collected from 42 Standardbred racehorses at rest and 60 min after performing a standardised treadmill exercise test. Tracheal wash and bronchoalveolar lavage fluid (BALF) samples were collected after exercise. Based on BALF cytology, 22 horses were defined as IAD‐affected and 20 classified as controls. Serum SP‐D concentrations were assessed using a commercially available ELISA kit and statistically compared between groups of horses and sampling times.

Results:Serum concentrations of SP‐D in IAD‐affected horses were significantly higher than those of control horses, both at rest and after exercise. Within the IAD‐affected group, no significant correlation was found between serum SP‐D concentrations and BALF cytology. Within each group of horses (IAD and control), no significant influence of exercise was found on serum SP‐D levels.

Conclusions:This is the first study determining serum SP‐D concentrations in a noninfectious, naturally occurring form of lower airway inflammation in horses. The results highlight that IAD is associated with a detectable, though moderate, increase of circulating SP‐D levels.

Potential relevance:Serum concentration of surfactant protein D could represent a potentially valuable and readily accessible blood biomarker of equine lower airway inflammation.
Retrospective analysis of DATIX dispensing error reports from Scottish NHS hospitals
Authors: Irwin Amy , Ross Jennifer , Seaton Janet , Mearns Kathryn
Published: June 2011

Aim

The primary objective was to analyse reported dispensing errors, and contributing factors, in Scottish National Health Service hospitals by coding and quantifying error reports from the DATIX patient-safety software. The secondary objective was to gather managerial responses to dispensing error in order to gain a perspective on interventions already in place.

Methods

Incident reports collected from 23 Scottish hospitals over a 5-year period were analysed retrospectively. Reported incident types, contributory factors and managerial responses were categorised according to the event description, and the frequency of such factors calculated.

Key findings

Dispensing errors (n = 573), from both pharmacies and wards, were analysed. The main incident types were incorrect drug (19.2%, n = 110) and incorrect strength of drug (16.8%, n = 96). The main contributory factors were reported as drug name similarity (15.5%, n = 30) and busy wards/pharmacies (14.9%, n = 29). Patient-centred issues (6.1%, n = 12) also featured. Managerial responses to these errors took the form of meetings (16.7%, n = 42), increasing staff awareness (14.7%, n = 37) or staff reminders on the importance of checking procedures (17.9%, n = 45).

Conclusions

The pattern of incidents reported is similar to previous research on the subject, but with a few key differences, such as, reports of errors associated with filling dosette boxes, and patient-centred issues. These differences indicate a potentially changing pattern of errors in response to new techniques in medicine management. Continued assessment of dispensing errors is required in order to develop practical interventions to improve medication safety.
Monoclonal Antibodies Specific for Immunorecessive Epitopes of Glucuronoxylomannan, the Major Capsular Polysaccharide of Cryptococcus neoformans, Reduce Serotype Bias in an Immunoassay for Cryptococcal Antigen
Authors: Percival Ann , Thorkildson Peter , Kozel Thomas R.
Published: June 2011

Immunoassay for detection of glucuronoxylomannan (GXM), the major capsular polysaccharide ofCryptococcus neoformans, is an important tool for diagnosis of cryptococcosis. However, immunoassays that are based solely or in part on detection with polyclonal antibodies may show serotype bias in detection of GXM, particularly limited sensitivity for serotype C. In this study, we describe detection of GXM in an antigen capture sandwich enzyme-linked immunosorbent assay (ELISA) that used a cocktail of two monoclonal antibodies (MAbs). MAb F12D2 was previously produced by immunization with GXM that had been treated to removeO-acetyl groups, a major source of serotype specificity. MAb F12D2 has a high degree of reactivity with GXM of serotypes A, B, C, and D, but the reactivity with serotype D was less than was found with other MAbs. MAb 339 is highly reactive with GXM of serotypes A and D. Use of a combination of the two MAbs produced an immunoassay that had the best properties of both MAbs, including good reactivity with serotype C, which is an emerging threat in sub-Saharan Africa. These results suggest that next-generation immunoassays for diagnosis of cryptococcosis may be formulated by (i) use of immunization and hybridoma screening strategies that are designed to prospectively meet the needs of immunoassay performance and (ii) careful selection of MAbs that span the expected polysaccharide serotypes in the subject patient population.
A Novel Lawsonia intracellularis Autotransporter Protein Is a Prominent Antigen
Authors: Watson Eleanor , Clark Ewan M. , Alberdi M. Pilar , Inglis Neil F. , Porter Megan , Imrie Lisa , Mclean Kevin , Manson Erin , Lainson Alex , Smith David G. E.
Published: June 2011

Investigation of antigenic determinants of the microaerophilic obligate intracellular bacteriumLawsonia intracellularisusing a mass spectrometry approach identified a novel bacterial protein present in an extract of cell culture medium aspirated from heavily infectedin vitrocell cultures. Western immunoblotting analysis of SDS-PAGE-resolved proteins using immune sera pooled fromL. intracellularis-infected pigs revealed the presence of a strongly immunoreactive band of ∼72 kDa. Liquid chromatography-electrospray ionization-tandem mass spectrometry analysis of this component and database mining using a fully annotatedL. intracellularisgenome sequence and the comprehensive GenBank prokaryotic genomic database highlighted the presence of a protein that shares little sequence similarity with other prokaryotic proteins and appears to be highly species specific. Detailed bioinformatic analyses identified the protein as member of the autotransporter protein family of surface-exposed proteins, and the designation LatA (Lawsoniaautotransporter protein A) is suggested. Recognition of recombinant LatA on Western blots by a panel of sera from infected and control pigs corresponded 100% with a commercial serodiagnostic that relies onin vitroculture of this fastidious organism. LatA therefore represents a potential candidate for the development of a rapid and species-specific serodiagnostic reagent.
Comparison of Memory B Cell, Antibody-Secreting Cell, and Plasma Antibody Responses in Young Children, Older Children, and Adults with Infection Caused by Vibrio cholerae O1 El Tor Ogawa in Bangladesh
Authors: Leung Daniel T. , Rahman Mohammad Arif , Mohasin M. , Riyadh M. Asrafuzzaman , Patel Sweta M. , Alam Mohammad Murshid , Chowdhury Fahima , Khan Ashraful Islam , Kalivoda Eric J. , Aktar Amena , Bhuiyan M. Saruar , LaRocque Regina C. , Harris Jason B. , Calderwood Stephen B. , Qadri Firdausi , Ryan Edward T.
Published: June 2011

Children bear a large component of the global burden of cholera. Despite this, little is known about immune responses to cholera in children, especially those under 5 years of age. Cholera vaccine studies have demonstrated lower long-term protective efficacy in young children than in older children and adults. Memory B cell (MBC) responses may correlate with duration of protection following infection and vaccination. Here we report a comparison of immune responses in young children (3 to 5 years of age;n= 17), older children (6 to 17 years of age;n= 17), and adults (18 to 60 years of age;n= 68) hospitalized with cholera in Dhaka, Bangladesh. We found that young children had lower baseline vibriocidal antibody titers and higher fold increases in titer between day 2 and day 7 than adults. Young children had higher baseline IgG plasma antibody levels toVibrio choleraeantigens, although the magnitudes of responses at days 7 and 30 were similar across age groups. As a surrogate marker for mucosal immune responses, we assessed day 7 antibody-secreting cell (ASC) responses. These were comparable across age groups, although there was a trend for older age groups to have higher levels of lipopolysaccharide-specific IgA ASC responses. All age groups developed comparable MBC responses toV. choleraelipopolysaccharide and cholera toxin B subunit at day 30. These findings suggest that young children are able to mount robust vibriocidal, plasma antibody, ASC, and MBC responses againstV. choleraeO1, suggesting that under an optimal vaccination strategy, young children could achieve protective efficacy comparable to that induced in adults.
Group B Streptococci Causing Neonatal Infections in Barcelona Are a Stable Clonal Population: 18-Year Surveillance
Authors: Martins E. R. , Andreu A. , Correia P. , Juncosa T. , Bosch J. , Ramirez M. , Melo-Cristino J.
Published: June 2011

We analyzed 212 group B streptococci (GBS) from newborns with invasive infections in the area of Barcelona, Spain, between 1992 and 2009, with the aim of documenting changes in the prevalences of serotypes, antimicrobial resistance, and genetic lineages and evaluating their associations with either early-onset disease (EOD) or late-onset disease (LOD). Serotypes III (
n
= 118) and Ia (
n
= 47) together accounted for nearly 78% of the isolates. All isolates carried an alpha or alpha-like protein gene, and specific associations between genes and serotypes, such as serotype Ib and
bca
, serotype II and
bca
, serotype III and
rib
, and serotype V and
alp3
, reflected the presence of particular genetic lineages. Macrolide resistance (14.2%) was significantly associated with serotype V. Pulsed-field gel electrophoresis (PFGE) clustering was an excellent predictor of serotype and antibiotic resistance. The combination of PFGE and multilocus sequence typing revealed a large number of genetically distinct lineages. Still, specific lineages were dominant in our collection, particularly the serotype III/ST17/
rib
lineage, which had enhanced potential to cause LOD. Serotype Ia was concentrated in a single PFGE cluster composed of two genetic lineages: ST23/
eps
and ST24/
bca
. The ST24/
bca
sublineage of serotype Ia, which is found infrequently elsewhere, may be emerging as an important cause of neonatal invasive infections in the Mediterranean region. In spite of the introduction of prophylaxis, resulting in a pronounced decline in the frequency of EOD, the study revealed a remarkably stable clonal structure of GBS causing neonatal infections in Barcelona over a period of 18 years.
Multilaboratory Evaluation of Real-Time PCR Tests for Hepatitis B Virus DNA Quantification
Authors: Caliendo Angela M. , Valsamakis Alexander , Bremer James W. , Ferreira-Gonzalez Andrea , Granger Suzanne , Sabatini Linda , Tsongalis Gregory J. , Wang Yun F. (Wayne) , Yen-Lieberman Belinda , Young Steve , Lurain Nell S.
Published: June 2011

The performance characteristics of four different assays for hepatitis B virus (HBV) quantification were assessed: the Abbott RealTime HBV IUO, the Roche Cobas AmpliPrep/Cobas TaqMan HBV test, the Roche Cobas TaqMan HBV test with HighPure system, and the Qiagen
artus
HBV TM ASR. Limit of detection (LOD), linear range, reproducibility, and agreement were determined using a serially diluted plasma sample from a single chronically infected subject. Each assay was tested by at least three laboratories. The LOD of the RealTime and two TaqMan assays was approximately 1.0 log
10
IU/ml; for
artus
HBV (which used the lowest volume of extracted DNA), it was approximately 1.5 log
10
IU/ml. The linear range spanned 1.0 to at least 7.0 log
10
IU/ml for all assays. Median values were consistently lowest for
artus
HBV and highest for Cobas AmpliPrep/Cobas TaqMan HBV. Assays incorporating automated nucleic acid extraction were the most reproducible; however, the overall variability was minor since the standard deviations for the means of all tested concentrations were ≤0.32 log
10
IU/ml for all assays. False-positive results were observed with all assays; the highest rates occurred with tests using manual nucleic acid extraction. The performance characteristics of these assays suggest that they are useful for management and therapeutic monitoring of chronic HBV infection.
Aeromonas aquariorum Is Widely Distributed in Clinical and Environmental Specimens and Can Be Misidentified as Aeromonas hydrophila
Authors: Aravena-Román Max , Harnett Gerald B. , Riley Thomas V. , Inglis Timothy J. J. , Chang Barbara J.
Published: June 2011

Genotypic characterization of 215
Aeromonas
strains (143 clinical, 52 environmental, and 20 reference strains) showed that
Aeromonas aquariorum
(60 strains, 30.4%) was the most frequently isolated species in clinical and water samples and could be misidentified as
Aeromonas hydrophila
by phenotypic methods.
Multiplex Real-Time PCR Assay for Detection of Methicillin-Resistant Staphylococcus aureus and Associated Toxin Genes
Authors: Fosheim G. E. , Nicholson A. C. , Albrecht V. S. , Limbago B. M.
Published: June 2011

We describe a real-time PCR assay for the detection of methicillin-resistant
Staphylococcus aureus
and genes encoding toxic shock syndrome toxin 1 and Panton-Valentine leukocidin. Rapid screening and detection of toxins is a useful tool for surveillance studies and outbreak investigations involving large numbers of isolates.
Development and Application of Hepatitis C Reporter Viruses with Genotype 1 to 7 Core-Nonstructural Protein 2 (NS2) Expressing Fluorescent Proteins or Luciferase in Modified JFH1 NS5A
Authors: Gottwein Judith M. , Jensen Tanja B. , Mathiesen Christian K. , Meuleman Philip , Serre Stephanie B. N. , Lademann Jacob B. , Ghanem Lubna , Scheel Troels K. H. , Leroux-Roels Geert , Bukh Jens
Published: June 2011

To facilitate genotype-specific high-throughput studies of hepatitis C virus (HCV), we have developed reporter viruses using JFH1-based recombinants expressing core-nonstructural protein 2 (NS2) of genotype 1 to 7 prototype isolates. We introduced enhanced green fluorescent protein (EGFP) into NS5A domain III of the genotype 2a virus J6/JFH1 [2a(J6)]. During Huh7.5 cell culture adaptation, 2a(J6)-EGFP acquired a 40-amino-acid (aa) (Δ40) or 25-aa (Δ25) deletion in NS5A domain II, rescuing the impairment of viral assembly caused by the EGFP insertion. Δ40 conferred efficient growth characteristics to 2a(J6) tagged with EGFP, DsRed-Express2, mCherry, or
Renilla
luciferase (RLuc), yielding peak supernatant infectivity titers of 4 to 5 log
10
focus-forming units (FFU)/ml. 2a(J6) with Δ40 or Δ25 was fully viable in Huh7.5 cells. In human liver chimeric mice, 2a(J6)-EGFPΔ40 acquired various deletions in EGFP, while 2a(J6)Δ40 did not show an impaired viability. We further developed panels of JFH1-based genotype 1 to 7 core-NS2 recombinants expressing EGFP- or RLuc-NS5AΔ40 fusion proteins. In cell culture, the different EGFP recombinants showed growth characteristics comparable to those of the nontagged recombinants, with peak infectivity titers of 4 to 5 log
10
FFU/ml. RLuc recombinants showed slightly less efficient growth characteristics, with peak infectivity titers up to 10-fold lower. Overall, the EGFP and RLuc recombinants were genetically stable after one viral passage. The usefulness of these reporter viruses for high-throughput fluorescence- and luminescence-based studies of HCV-receptor interactions and serum-neutralizing antibodies was demonstrated. Finally, using RLuc viruses, we showed that the genotype-specific core-NS2 sequence did not influence the response to alfa-2b interferon (IFN-alfa-2b) and that genotype 1 to 7 viruses all responded to treatment with p7 ion channel inhibitors.
Potyviral VPg Enhances Viral RNA Translation and Inhibits Reporter mRNA Translation
In Planta
Authors: Eskelin Katri , Hafrén Anders , Rantalainen Kimmo I. , Mäkinen Kristiina
Published: June 2011

Viral protein genome-linked (VPg) plays a central role in several stages of potyvirus infection. This study sought to answer questions about the role of
Potato virus A
(PVA; genus
Potyvirus
) VPg in viral and host RNA expression. When expressed in
Nicotiana benthamiana
leaves
in trans
, a dual role of VPg in translation is observed. It repressed the expression of monocistronic luciferase (
luc
) mRNA and simultaneously induced a significant upregulation in the expression of both replicating and nonreplicating PVA RNAs. This enhanced viral gene expression was due at least to the 5′ untranslated region (UTR) of PVA RNA, eukaryotic initiation factors 4E and iso 4E [eIF4E/eIF(iso)4E], and the presence of a sufficient amount of VPg. Coexpression of VPg with viral RNA increased the viral RNA amount, which was not the case with the monocistronic mRNA. Both mutations at certain lysine residues in PVA VPg and eIF4E/eIF(iso)4E depletion reduced its ability to upregulate the viral RNA expression. These modifications were also involved in VPg-mediated downregulation of monocistronic
luc
expression. These results suggest that VPg can titrate eIF4Es from capped monocistronic RNAs. Because VPg-mediated enhancement of viral gene expression required eIF4Es, it is possible that VPg directs eIF4Es to promote viral RNA expression. From this study it is evident that VPg can serve as a specific regulator of PVA expression by boosting the viral RNA amounts as well as the accumulation of viral translation products. Such a mechanism could function to protect viral RNA from being degraded and to secure efficient production of coat protein (CP) for virion formation.
Type I Interferon Contributes to CD4+T Cell Depletion Induced by Infection with HIV-1 in the Human Thymus
Authors: Sivaraman Vijay , Zhang Liguo , Su Lishan
Published: June 2011

Persistent induction of type 1 interferon (IFN) is associated with human immunodeficiency virus type 1 (HIV-1) infection. We report here that the pathogenic HIV strain R3A (HIV-R3A) induced high levels of type 1 IFN, while the nonpathogenic HIV-R3B showed no significant induction in human fetal thymus organ culture (HFTOC). We found that IFN contributed to the depletion of human T cells by HIV-R3A in a fusion-independent fashion. The R3B recombinant with the R3A Env V1V2 domain (R3B/A-V1V2) was able to induce type 1 IFN, which contributed to the increased depletion of T cells. Therefore, type 1 IFN induction plays a significant role in HIV-induced T cell depletion in the human thymus.
APOBEC3 Has Not Left an Evolutionary Footprint on the HIV-1 Genome
Authors: Ebrahimi Diako , Anwar Firoz , Davenport Miles P.
Published: June 2011

It is known that the human immune proteins APOBEC3G and -F (hA3G/F) can inhibit Vif-deficient HIV by G-to-A mutation; however, the roles of these enzymes in the evolution of HIV are debated. We argue that if evolutionary pressure from hA3G/F exists there should be evidence of their imprint on the HIV genome in the form of (i) underrepresentation of hA3G/F target motifs (e.g., T
G
GG [targeted position is underlined]) and overrepresentation of product motifs (e.g., T
A
GG) and/or (ii) an increase in the ratio of nonsynonymous to synonymous (NS/S) G-to-A changes among hA3G/F target motifs and a decrease of NS/S A-to-G changes among hA3G/F product motifs. To test the first hypothesis, we studied the representation of hA3G/F target and product motifs in 1,932 complete HIV-1 genomes using Markov models. We found that the highly targeted motifs are not underrepresented and their product motifs are not overrepresented. To test the second hypothesis, we determined the NS/S G↔A changes among the hA3G/F target and product motifs in 1,540 complete sets of nine HIV-1 genes. The NS/S changes did not show an increasing/decreasing trend within the target/product motifs, but the NS/S changes within the motif
A
G was exceptionally low. We observed the same pattern by analyzing 740 human genes. Given that hA3G/F do not act on the human genome, this suggests a small NS/S change within
A
G has arisen by other mechanisms. We therefore find no evidence of an evolutionary footprint of hA3G/F. We postulate several mechanisms to explain why the HIV-1 genome does not contain the hA3G/F footprint.
Direct Inhibition of Tombusvirus Plus-Strand RNA Synthesis by a Dominant Negative Mutant of a Host Metabolic Enzyme, Glyceraldehyde-3-Phosphate Dehydrogenase, in Yeast and Plants
Authors: Huang Tyng-Shyan , Nagy Peter D.
Published: June 2011

The replication of plus-strand RNA viruses depends on many cellular factors. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an abundant metabolic enzyme that is recruited to the replicase complex of
Tomato bushy stunt virus
(TBSV) and affects asymmetric viral RNA synthesis. To further our understanding on the role of GAPDH in TBSV replication, we used an
in vitro
TBSV replication assay based on recombinant p33 and p92
pol
viral replication proteins and cell-free yeast extract. We found that the addition of purified recombinant GAPDH to the cell extract prepared from GAPDH-depleted yeast results in increased plus-strand RNA synthesis and asymmetric production of viral RNAs. Our data also demonstrate that GAPDH interacts with p92
pol
viral replication protein, which may facilitate the recruitment of GAPDH into the viral replicase complex in the yeast model host. In addition, we have identified a dominant negative mutant of GAPDH, which inhibits RNA synthesis and RNA recruitment
in vitro
. Moreover, this mutant also exhibits strong suppression of tombusvirus accumulation in yeast and in virus-infected
Nicotiana benthamiana
. Overall, the obtained data support the model that the co-opted GAPDH plays a direct role in TBSV replication by stimulating plus-strand synthesis by the viral replicase.
Kaposi's Sarcoma-Associated Herpesvirus-Encoded Latency-Associated Nuclear Antigen Reduces Interleukin-8 Expression in Endothelial Cells and Impairs Neutrophil Chemotaxis by Degrading Nuclear p65
Authors: Li Xiaofan , Liang Deguang , Lin Xianzhi , Robertson Erle S. , Lan Ke
Published: June 2011

Latency-associated nuclear antigen 1 (LANA-1) of Kaposi's sarcoma-associated herpesvirus (KSHV) is the major viral latent protein and functions as a multifaceted protein. Here, we report that LANA-1 attenuates the endothelial response to tumor necrosis factor alpha (TNF-α) stimulation and inhibits consequent neutrophil chemotaxis. Reporter assays showed that LANA-1 constantly repressed nuclear factor (NF)-κB transactivity upon TNF-α stimulation. We also found that LANA-1 decreased nuclear p65 protein levels through enhancement of polyubiquitylation-mediated p65 degradation and that an elongin B/elongin C–cullin 5–LANA-1–p65 complex assembled by LANA-1 was responsible for this enhanced p65 degradation. In telomerase-immortalized human umbilical vein endothelial cells, LANA-1 was demonstrated to repress interleukin-8 expression, which was involved in neutrophil recruitment to the inflammatory site. Through anin vitrotransmigration assay, we determined a suppressive effect of LANA-1 on neutrophil chemotaxis. Our work suggests that KSHV LANA-1 is a negative modulator of acute inflammation and sheds light on a new mechanism by which KSHV during the latent life cycle evades the host innate immune response.
Characterization of H1N1 Swine Influenza Viruses Circulating in Canadian Pigs in 2009
Authors: Nfon Charles K. , Berhane Yohannes , Hisanaga Tamiko , Zhang Shunzhen , Handel Katherine , Kehler Helen , Labrecque Olivia , Lewis Nicola S. , Vincent Amy L. , Copps John , Alexandersen Soren , Pasick John
Published: June 2011

The 2009 pandemic H1N1 (pH1N1), of apparent swine origin, may have evolved in pigs unnoticed because of insufficient surveillance. Consequently, the need for surveillance of influenza viruses circulating in pigs has received added attention. In this study we characterized H1N1 viruses isolated from Canadian pigs in 2009. Isolates from May 2009 were comprised of hemagglutinin and neuraminidase (NA) genes of classical SIV origin in combination with the North American triple-reassortant internal gene (TRIG) cassette, here termed contemporary SIV (conSIV) H1N1. These conSIV H1N1 viruses were contiguous with the North American αH1 cluster, which was distinct from the pH1N1 isolates that were antigenically more related to the γH1 cluster. After the initial isolation of pH1N1 from an Alberta pig farm in early May 2009, pH1N1 was found several times in Canadian pigs. These pH1N1 isolates were genetically and antigenically homogeneous. In addition, H1N1 viruses bearing seasonal human H1 and N1 genes together with the TRIG cassette and an NA encoding an oseltamivir-resistance marker were isolated from pigs. The NS gene of one of these seasonal human-like SIV (shSIV) H1N1 isolates was homologous to pH1N1 NS, implicating reassortment between the two strains. Antigenic cross-reactivity was observed between pH1N1 and conSIV but not with shSIV H1N1. In summary, although there was cocirculation of pH1N1 with conSIV and shSIV H1N1 in Canadian pigs after May 2009, there was no evidence supporting the presence of pH1N1 in pigs prior to May 2009. The possibility for further reassortants being generated exists and should be closely monitored.
Complete Genome Analysis of Three Novel Picornaviruses from Diverse Bat Species
Authors: Lau Susanna K. P. , Woo Patrick C. Y. , Lai Kenneth K. Y. , Huang Yi , Yip Cyril C. Y. , Shek Chung-Tong , Lee Paul , Lam Carol S. F. , Chan Kwok-Hung , Yuen Kwok-Yung
Published: June 2011

Although bats are important reservoirs of diverse viruses that can cause human epidemics, little is known about the presence of picornaviruses in these flying mammals. Among 1,108 bats of 18 species studied, three novel picornaviruses (groups 1, 2, and 3) were identified from alimentary specimens of 12 bats from five species and four genera. Two complete genomes, each from the three picornaviruses, were sequenced. Phylogenetic analysis showed that they fell into three distinct clusters in the
Picornaviridae
family, with low homologies to known picornaviruses, especially in leader and 2A proteins. Moreover, group 1 and 2 viruses are more closely related to each other than to group 3 viruses, which exhibit genome features distinct from those of the former two virus groups. In particular, the group 3 virus genome contains the shortest leader protein within
Picornaviridae
, a putative type I internal ribosome entry site (IRES) in the 5′-untranslated region instead of the type IV IRES found in group 1 and 2 viruses, one instead of two GXCG motifs in 2A, an L→V substitution in the DDLXQ motif in 2C helicase, and a conserved GXH motif in 3C protease. Group 1 and 2 viruses are unique among picornaviruses in having AMH instead of the GXH motif in 3C
pro
. These findings suggest that the three picornaviruses belong to two novel genera in the
Picornaviridae
family. This report describes the discovery and complete genome analysis of three picornaviruses in bats, and their presence in diverse bat genera/species suggests the ability to cross the species barrier.
The Rhesus Rotavirus Gene Encoding VP4 Is a Major Determinant in the Pathogenesis of Biliary Atresia in Newborn Mice
Authors: Wang Wei , Donnelly Bryan , Bondoc Alexander , Mohanty Sujit K. , McNeal Monica , Ward Richard , Sestak Karol , Zheng Shan , Tiao Greg
Published: June 2011

Biliary atresia (BA) is a devastating disease of childhood for which increasing evidence supports a viral component in pathogenesis. The murine model of BA is induced by perinatal infection with rhesus rotavirus (RRV) but not with other strains of rotavirus, such as TUCH. To determine which RRV gene segment(s) is responsible for pathogenesis, we used the RRV and TUCH strains to generate a complete set of single-gene reassortants. Eleven single-gene “loss-of-function” reassortants in which a TUCH gene replaced its RRV equivalent and 11 single-gene “gain-of-function” reassortants in which an RRV gene replaced its TUCH equivalent were generated. Newborn BALB/c mice were inoculated with the reassortants and were monitored for biliary obstruction and mortality.
In vitro
, the ability to bind to and replicate within cholangiocytes was analyzed. Infection of mice with the “loss-of-function” reassortant R
T(VP4)
, where gene 4 from TUCH was placed on an RRV background, eliminated the ability of RRV to cause murine BA. In a reciprocal fashion, the “gain-of-function” reassortant T
R(VP4)
resulted in murine BA with 88% mortality. Compared with those for RRV, R
T(VP4)
binding and titers in cholangiocytes were significantly attenuated, while T
R(VP4)
binding and titers were significantly increased over those for TUCH. Reassortants R
T(VP3)
and T
R(VP3)
induced an intermediate phenotype. RRV gene segment 4 plays a significant role in governing tropism for the cholangiocyte and the ability to induce murine BA. Gene segment 3 did not affect RRV infectivity
in vitro
but altered its
in vivo
effect.
Binding of Avian Coronavirus Spike Proteins to Host Factors Reflects Virus Tropism and Pathogenicity
Authors: Wickramasinghe I. N. Ambepitiya , de Vries R. P. , Gröne A. , de Haan C. A. M. , Verheije M. H.
Published: June 2011

The binding of viruses to host cells is the first step in determining tropism and pathogenicity. While avian infectious bronchitis coronavirus (IBV) infection and avian influenza A virus (IAV) infection both depend on α2,3-linked sialic acids, the host tropism of IBV is restricted compared to that of IAV. Here we investigated whether the interaction between the viral attachment proteins and the host could explain these differences by using recombinant spike domains (S1) of IBV strains with different pathogenicities, as well as the hemagglutinin (HA) protein of IAV H5N1. Protein histochemistry showed that S1 of IBV strain M41 and HA of IAV subtype H5N1 displayed sialic acid-dependent binding to chicken respiratory tract tissue. However, while HA bound with high avidity to a broad range of α2,3-linked sialylated glycans, M41 S1 recognized only one particular α2,3-linked disialoside in a glycan array. When comparing the binding of recombinant IBV S1 proteins derived from IBV strains with known differences in tissue tropism and pathogenicity, we observed that while M41 S1 displayed binding to cilia and goblet cells of the chicken respiratory tract, S1 derived from the vaccine strain H120 or the nonvirulent Beaudette strain had reduced or no binding to chicken tissues, respectively, in agreement with the reduced abilities of these viruses to replicate
in vivo
. While the S1 protein derived from the nephropathogenic IBV strain B1648 also hardly displayed binding to respiratory tract cells, distinct binding to kidney cells was observed, but only after the removal of sialic acid from S1. In conclusion, our data demonstrate that the attachment patterns of the IBV S proteins correlate with the tropisms and pathogenicities of the corresponding viruses.
The Left Half of the XMRV Retrovirus Is Present in an Endogenous Retrovirus of NIH/3T3 Swiss Mouse Cells
Authors: Mendoza Ramon , Vaughan Andrew E. , Miller A. Dusty
Published: June 2011

Xenotropic murine leukemia virus-related virus (XMRV) is a gammaretrovirus found in association with human prostate cancer and chronic fatigue syndrome, although these associations are controversial. XMRV shows at most 94% identity to known mouse retroviruses. Here we used XMRV-specific PCR to search for a more closely related source of XMRV in mice. While we could not find a complete copy, we did find a 3,600-bp region of XMRV in an endogenous retrovirus present in NIH/3T3 cells. These results show that XMRV has clear ancestors in mice and highlight another possible source of contamination in PCR assays for XMRV.
Enhanced Sialic Acid-Dependent Endocytosis Explains the Increased Efficiency of Infection of Airway Epithelia by a Novel Adeno-Associated Virus
Authors: Dickey David D. , Excoffon Katherine J. D. A. , Koerber James T. , Bergen Jamie , Steines Benjamin , Klesney-Tait Julia , Schaffer David V. , Zabner Joseph
Published: June 2011

We previously used directed evolution in human airway epithelia to create adeno-associated virus 2.5T (AAV2.5T), a highly infectious chimera of AAV2 and AAV5 with one point mutation (A581T). We hypothesized that the mechanism for its increased infection may be a higher binding affinity to the surface of airway epithelia than its parent AAV5. Here, we show that, like AAV5, AAV2.5T, uses 2,3N-linked sialic acid as its primary receptor; however, AAV2.5T binds to the apical surface of human airway epithelia at higher levels and has more receptors than AAV5. Furthermore, its binding affinity is similar to that of AAV5. An alternative hypothesis is that AAV2.5T interaction with 2,3N-linked sialic acid may instead be required for cellular internalization. Consistent with this, AAV2.5T binds but fails to be internalized by CHO cells that lack surface expression of sialic acid. Moreover, whereas AAV2.5T binds similarly to human (rich in 2,3N-linked sialic acid) and pig airway epithelia (2,6N-linked sialic acid), significantly more virus was internalized by human airway. Subsequent transduction correlated with the level of internalized rather than surface-bound virus. We also found that human airway epithelia internalized significantly more AAV2.5T than AAV5. These data suggest that AAV2.5T has evolved to utilize specific 2,3N-linked sialic acid residues on the surface of airway epithelia that mediate rapid internalization and subsequent infection. Thus, sialic acid serves as not just an attachment factor but is also required for AAV2.5T internalization, possibly representing an important rate-limiting step for other viruses that use sialic acids.
Is the commercialisation of human tissue and body material forbidden in the countries of the European Union?
Authors: Lenk Christian , Beier Katharina
Published: June 2011

The human body and its parts are widely perceived as matters beyond commercial usage. This belief is codified in several national and European documents. This so-called ‘no-property rule’ is held to be the default position across the countries of the European Union. However, a closer look at the most pertinent national and European documents, and also current practices in the field, reveals a gradual model of commercialisation of human tissue. In particular, we will argue that the ban on commercialisation of body material is not as strict as it may appear at first sight, leaving room for the commercial practice of tissue procurement and transfer. We argue for more transparent information for patients and tissue donors, an intensified ethical debate on commercialisation practices, and a critical review of current normative principles.
Clinical veterinary students' perceptions of a ‘Day one’ skills guide
Authors: Duncan C. , Dale V. H. M. , Pead M. J.
Published: June 2011

Veterinary schools in the UK are required by the Royal College of Veterinary Surgeons to ensure that their students have attained essential ‘Day one’ skills (DOS) by graduation. To aid students' understanding of how they could satisfy this requirement, the Royal Veterinary College produced a DOS guide, which was distributed in the 2007 to 2008 academic year to students in the third and fourth years of the professional veterinary course. The same students were surveyed towards the end of the 2008 to 2009 session about the guide, when they were in the fourth and final years, to rate its usefulness and to indicate their perceived levels of competence, both currently and expected at graduation. Focus group discussions were subsequently conducted to explore the findings of the survey. Responses from 88 fourth‐year (39 per cent) and 174 (87 per cent) final‐year students revealed that while almost all respondents were aware of the DOS guide, their use of it was low. Final‐year students rated themselves as being more competent than fourth‐year students, but were less optimistic about their expected level of competence regarding several invasive procedures, which some students considered to be ‘unattainable’ by graduation, despite the apparent value of extramural studies and intramural rotations in providing opportunities for learning.
MULTI-STATE START-UP DEMONSTRATION TESTS
Authors: SMITH MICHELLE L. DEPOY , GRIFFITH WILLIAM S.
Published: June 2011

Multistate start-up demonstration tests are proposed and analyzed using a Markov Chain approach. The new demonstration tests are generalizations of the CSTF, TSTF, and CSCF start-up demonstration tests discussed in the literature that have the important practical advantage of allowing for early termination when the quality of successful start-ups is exceptional. When the termination results in acceptance of the equipment, the use of a multi-state startup demonstration test results in an increase in the probability of an early termination of as much as. 35 over the optimal binary start-up test in cases studied. The Markov Chains methodology is used in the probabilistic analysis. Practical guidance on choosing tests, estimation, and comparisons of the various criteria are studied. The Markov Chain approach is then extended to study the non-i.i.d. case.