Analysis of bovine viral diarrhoea virus: Biobank and sequence database to support eradication in Scotland.

Samples from bovine viral diarrhoea virus (BVDV)-positive cattle were gathered by Scottish diagnostic laboratories and used to produce a Biobank of samples with associated location and identification data in support of the Scottish BVDV eradication scheme. The samples were subject to direct amplification and sequencing of the 5'-untranslated region (5'-UTR) to define the viral types and subtypes present. From 2693 samples collected prior to 2016, approximately 2300 sequences were obtained, representing 8 BVDV type 1 subtypes. No BVDV type 2 samples were detected. The samples came from all regions of the UK but 66 per cent were from Scotland. Analysis of the sequences showed great diversity in the 5'-UTR, with 1206 different sequences. Many samples carried virus with identical 5'-UTR sequences; often from single locations, but there were also examples of the same sequence being obtained from samples at several different locations. This work provides a resource that can be used to analyse the movement of BVDV strains both within Scotland and between Scotland and other nations, particularly in the latter stages of the Scottish eradication programme, and so inform the advice available to both livestock keepers and policymakers.

Development of Xe and Kr empirical potentials for CeO2, ThO2, UO2 and PuO2, combining DFT with high temperature MD.

The development of embedded atom method (EAM) many-body potentials for actinide oxides and associated mixed oxide (MOX) systems has motivated the development of a complementary parameter set for gas-actinide and gas-oxygen interactions. A comprehensive set of density functional theory (DFT) calculations were used to study Xe and Kr incorporation at a number of sites in CeO2, ThO2, UO2 and PuO2. These structures were used to fit a potential, which was used to generate molecular dynamics (MD) configurations incorporating Xe and Kr at 300 K, 1500 K, 3000 K and 5000 K. Subsequent matching to the forces predicted by DFT for these MD configurations was used to refine the potential set. This fitting approach ensured weighted fitting to configurations that are thermodynamically significant over a broad temperature range, while avoiding computationally expensive DFT-MD calculations. The resultant gas potentials were validated against DFT trapping energies and are suitable for simulating combinations of Xe and Kr in solid solutions of CeO2, ThO2, UO2 and PuO2, providing a powerful tool for the atomistic simulation of conventional nuclear reactor fuel UO2 as well as advanced MOX fuels.

[Fluorquinolone associated tendon rupture]. / Achillessehnenruptur unter Fluorochinolonen.

We report the case of a 38-year-old patient with a rupture of the right Achilles tendon after physical exercise. A few days before he had been treated with ciprofloxacine 500mg bid for chlamydial urethritis. We discuss know risk factors for Achilles tendon ruptures and the possible contribution of ciprofloxacin and fluorquinolones in this case.

The efficacy of a commercial ELISA as an alternative to virus neutralisation test for the detection of antibodies to EAV.

Infection with equine arteritis virus is a notifiable disease with sporadic occurrence in the UK. As stallions may harbour the virus after infection, horses are screened for exposure by serological testing prior to breeding. The virus neutralisation test is considered the 'gold standard' serological screening test, but it is time-consuming and labour intensive; consequently there is a move towards more rapid screening methodology. In this study, a commercially available EVA antibody ELISA is assessed. The ELISA performed poorly with a specificity [corrected] of 26% and a sensitivity [corrected] of 96% in the samples analysed. It was concluded that this ELISA would be of little value for reducing sample turnaround time. The study emphasises the need for in-house validation of commercially available kits.

Factors affecting the serological response of dogs and cats to rabies vaccination.

After being vaccinated against rabies some cats and dogs fail to show an antibody titre adequate to meet the requirements of the UK Pet Travel Scheme. To investigate this problem, the data derived from 16,073 serum samples submitted to the Veterinary Laboratories Agency for serological testing between 1999 and 2002, 1002 samples submitted to BioBest during March and April 2001, and 1264 samples associated with one make of vaccine submitted to BioBest between June 2001 and January 2003, were analysed. The probability of antibody titre failing to reach at least 0.5 iu/ml was analysed by logistic regression as a function of the choice of vaccine, the interval between vaccination and sampling, the sex and age of the animal, and its country of origin. In dogs, all these factors, except sex, had highly significant (P < 0.001) effects on the test failure rate, and in cats all the factors had a significant effect (P < 0.05).

Involvement of NMDA receptors and a p21Ras-like guanosine triphosphatase in the constitutive activation of nuclear factor-kappa-B in cortical neurons.

The transcription factor nuclear factor-kappa-B (NF-kappaB) is now recognised as a key mediator of physiological and pathological plasticity in the central nervous system (CNS), and ionotropic glutamate receptor stimulation potently triggers NF-kappaB activation. This study was designed to identify the mechanisms responsible for the high basal levels of activated NF-kappaB present in neurons in the cerebral cortex. In cultured cortical neurons, the basal levels of activated NF-kappaB were reduced by the glutamate receptor antagonists MK801 and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), but were not affected by exposure to a mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor, a p38 MAP kinase inhibitor or a cyclic guanosine monophosphate (cGMP)-dependent protein kinase inhibitor. However, activated NF-kappaB levels were reduced by a guanylate cyclase inhibitor, the Src-family tyrosine kinase inhibitor PP1, or the farnesyl transferase inhibitors manumycin and farnesyl transferase (Ftase) inhibitor 1. There was no additive effect when MK801 was applied together with manumycin. These results suggest that the basal levels of activated NF-kappaB in cortical neurons are maintained partially by synaptic activity involving N-methyl- D-aspartate (NMDA) and AMPA/kainate glutamate receptors, coupled to activation of an Src-family tyrosine kinase and a p21(Ras)-like guanosine triphosphatase (GTPase) in a cGMP-dependent manner. The results are intriguing in the light of the recent identification of a synaptic p21(Ras) activator stimulated by cGMP.

Cloning, sequence analysis and expression of the cDNAs encoding the canine and equine homologues of the mouse double minute 2 (mdm2) proto-oncogene.

The mdm2 oncogene is amplified and overexpressed in a variety of human tumours and the oncogenic potential of MDM2 is partly due to its ability to inactivate tumour suppressor p53 function. In the present communication we describe the cloning, sequence analysis and expression of the complete wildtype canine and equine mdm2 cDNAs. The encoded full-length canine and equine cDNAs show strong sequence homology with MDM2 proteins from other species and both cDNAs generate recombinant proteins of approximately 90 kDa. These data will allow for the role of this oncogene to be established in companion animal oncology.

Nucleotide sequence of the porcine p53 cDNA, and the detection of recombinant porcine p53 expressed in vitro with a variety of anti-p53 antibodies.

The cDNA of porcine p53 was cloned and sequenced by a reverse transcriptase polymerase chain reaction (RT - PCR) approach with primers based on regions of homology between all known p53 sequences. The p53 cDNA was found to be 87% conserved to human p53 and 86% conserved to bovine p53 at the nucleotide level. The porcine p53 sequence was inserted into an expression vector and recombinant protein expressed in vitro. An approximately 50 kDa protein was detected by Western blotting using both polyclonal and monoclonal anti-p53 antibodies. The sequence data of porcine p53 and the ability to detect expressed protein with various anti-p53 antibodies will allow the p53 status of the pig population, and the role of p53 in porcine tumours, to be assessed. An understanding of tumour development in the pig may be important if pig cells, tissues or organs are to be used in the treatment of humans.

Detection of canine herpesvirus 1 in a wide range of tissues using the polymerase chain reaction.

Canine herpesvirus 1 (CHV-1), a member of the alphaherpesvirus sub-family, is known to cause fatal infections in litters of puppies and may also be involved in infertility, abortion, and stillbirths in adult dogs. The purpose of this study was to determine the presence of CHV-1 DNA using the polymerase chain reaction (PCR) in twelve key sites that have been associated with latency for the other herpesviruses. A 605 base pair portion of the viral glycoprotein B (gB) gene was amplified using degenerate primers, cloned, and sequenced. Conventional 20 mer primers were designed using this sequence information to amplify a 120 bp fragment of gB situated between the original degenerate primers. The specificity of amplification was confirmed by Southern Blot hybridisation using an internal oligonucleotide probe. DNA was extracted from tissue samples taken from twelve dogs at post mortem and from twenty-four blood samples. Nine out of twelve dogs showed evidence of infection with CHV-1; the tissues most commonly affected were lumbo-sacral ganglia (5/12 dogs), tonsil (5/12), parotid salivary gland (4/9), and liver (4/9). No positive results were detected within the twenty-four blood samples. These results indicate that exposure to CHV-1 may be much more common than previously suggested.

Risk factors predicting exertional heat illness in male Marine Corps recruits.

A matched population-based case-control study was conducted on exertional heat illness (EHI) in male Marine Corps recruits in basic training at Parris Island, SC. Physical fitness and anthropometric measurements were obtained for 391 of 528 cases of EHI identified in this population during 1988-1992, and 1467 of 1725 controls matched to cases by initial training platoon. The risk for developing EHI increased with increase in body mass index (BMI = weight.height-2) as measured on arrival and with increase in time to complete a 1.5-mile run conducted during the first week. Recruits at highest risk for developing exertional heat illness had a BMI of 22 or more kg.m-2 and a 1.5-mile run-time for 12 of more minutes. These recruits had an eightfold higher risk for developing exertional heat illness during basic training when compared with those with BMI less than 22 kg.m-2 and 1.5-mile run-time under 10 min (P < 10(-6). Only one-fifth (18%) of male recruits met these criteria for high risk, but they accounted for nearly half (47%) of the exertional heat illness cases occurring during the 12-wk basic training course.

Exertional heat illness in Marine Corps recruit training.

BACKGROUND: Exertional heat illness (EHI) is a recurring problem in military training resulting in loss of manpower and training effectiveness. A scientific approach to this problem requires quantitative analysis of factors related to its occurrence. METHODS: Review of clinic and hospital records identified all cases of EHI occurring among 217,000 Marine Corps recruits (90% male, 80% age 18-20 yr) entering 12-week basic training at Parris Island, SC during 1982-1991. RESULTS: There were 1454 cases, 89% males and 11% females; 11% of male and no female cases were hospitalized. The majority (88%) of cases occurred during May-September, with a peak rate of 2% of recruits in summer. Most of the cases occurred during the cooler early morning hours when recruits performed strenuous exercise. During 0700-0900 hours the rate of heat casualties increased substantially as the Wet Bulb Globe Temperature Index (WBGT) increased, beginning at levels as low as 65 degrees F. EHI rates increased 26- and 39-fold at WBGT 75- < 80 degrees F (over baseline rates at < 65 degrees F) for immediate and prior day exposures, respectively. Exposure to WBGT of 80 + degrees F was infrequent (25%) among the early morning cases at the time of illness, while it was common (87%) on the day prior to illness, suggesting a lasting effect of heat exposure. CONCLUSIONS: Risk for EHI in military training increases markedly as WBGT levels rise above 65 degrees F; cases occur primarily with strenuous exercise (e.g., running); and heat stress exposure on prior days may be important.