We use both large and small animal models in our pre-clinical evaluation of gene transfer agents (GTAs) for cystic fibrosis (CF) gene therapy. Here, we report the use of a large animal model to assess three non-viral GTAs: 25 kDa-branched polyethyleneimine (PEI), the cationic liposome (GL67A) and compacted DNA nanoparticle formulated with polyethylene glycol-substituted lysine 30-mer. GTAs complexed with plasmids expressing human cystic fibrosis transmembrane conductance regulator (CFTR) complementary DNA were administered to the sheep lung (n¼8 per group) by aerosol. All GTAs gave evidence of gene transfer and expression 1 day after treatment. Vector-derived mRNA was expressed in lung tissues, including epithelial cell-enriched bronchial brushing samples, with median group values reaching 1-10% of endogenous CFTR mRNA levels. GL67A gave the highest levels of expression. Human CFTR protein was detected in small airway epithelial cells in some animals treated with GL67A (two out of eight) and PEI (one out of eight). Bronchoalveolar lavage neutrophilia, lung histology and elevated serum haptoglobin levels indicated that gene delivery was associated with mild local and systemic inflammation. Our conclusion was that GL67A was the best non-viral GTA currently available for aerosol delivery to the sheep lung, led to the selection of GL67A as our lead GTA for clinical trials in CF patients.

We consider the pair correlation functions of both the order parameter field and its square for phase ordering in the O(n) model with nonconserved order parameter, in spatial dimension 2 ≤ d ≤ 3 and spin dimension 1 ≤ n ≤ d. We calculate, in the scaling limit, the exact short-distance singularities of these correlation functions and compare these predictions to numerical simulations. Our results suggest that the scaling hypothesis does not hold for the d = 2 O(2) model.

The 'H NMR spectrum of chlorpromazine hydrochloride was fully assigned at 400 MHz Similarly, the "C NMR spectrum was assigned unambiguously using two-dimensional NMR. Measurements of chemical shift as a function of concentration in D,O showed appreciable changes of shift of both protons and carbons which were apparent even at solution concentrations two orders of magnitude lower than the critical micelle concentration (CMC). The relative magnitude of the shifts of the aromatic protons and carbons on dilution below the CMC were compatible with vertical stacking of the molecules in an off-set manner such that maximum overlap of the chlorinated rings occurred. Proton chemical shift data were interpreted using a stepwise association model to quantify the extent of association in the pre-CMC region.

SummaryBackgroundEquine recurrent uveitis (ERU) is a common cause of ocular pain and blindness in horses. Leptospira spp. have been commonly implicated in the pathophysiology of ERU in mainland Europe and the USA. No recent studies have been carried out in the UK, but Leptospira is reported not to be a major factor in the aetiology of ERU in the UK.ObjectivesTo establish the prevalence of Leptospira‐associated ERU in the UK and to identify the serovars involved in these cases; to compare serum vs. aqueous humour antibody levels in cases and controls in order to confirm the diagnosis of Leptospira‐associated ERU, and to assess the usefulness of serology alone as a confirmatory test for Leptospira‐associated ERU in the UK.Study designCase–control study.MethodsEyes enucleated for clinical reasons in ERU‐affected horses were collected. Blood and aqueous humour were obtained to determine antibody levels against a variety of Leptospira serovars and C‐values (aqueous humour value/serum value) were calculated. In addition, eyes, blood and aqueous humour were obtained from control cases for comparison. Histopathology was performed in all eyes to confirm uveitis in each case. Differences in seroprevalences between ERU and control cases and between Leptospira‐ and non‐Leptospira‐associated ERU cases were calculated.ResultsA total of 30 ERU and 43 control eyes were analysed. Of the ERU eyes, only two had a C‐value of >4 (prevalence of Leptospira‐associated uveitis: 6.7%). Serovars hardjo and javanica were detected. There was no difference in seroprevalence between horses with uveitis and control cases (65.5% and 41.9%, respectively; P = 0.11) or between Leptospira‐ and non‐Leptospira‐associated uveitis cases (100% and 63.0%, respectively; P = 0.52).Main limitationsThe study was limited by low case numbers. Eyes were presented at different stages of disease. The only test used to detect Leptospira was the microscopic agglutination test.Conclusions Leptospira‐associated ERU is uncommon in the UK. Serology alone may not help to definitively diagnose Leptospira‐associated uveitis in this country.

Contagious ovine digital dermatitis (CODD) is an infectious foot disease of sheep causing severe lameness. Diagnosis is currently made using broad anecdotal descriptions. The aim of this study was to systematically and formally describe the clinical presentation of the disease in terms of (1) a lesion grading system; (2) associated radiographic changes and (3) severity of associated lameness. A five-point lesion grading system was developed and applied to 908 sheep affected by CODD from six farms. Sheep with lesions typical of each grade were euthanased and their feet radiographed. Radiographic abnormalities including soft tissue and bony changes were evident in feet with lesions graded 2-5. In order to quantify the welfare impact of CODD, all the 908 sheep were locomotion scored. Five hundred and eighty-five (64.5% (95% CI 61.4% to 67.6%)) were lame. The locomotion score for affected sheep increased with worsening pathological changes. Once healing had begun the locomotion score decreased. In conclusion the five-point grading system may be used to clinically describe stages of CODD lesions. The radiographic changes revealed examples of deeper pathological changes and there was a strong association between the lesion grading system and locomotion score in affected sheep.

We describe a case of zinc toxicity in a 14-month-old, female, neutered, Cavalier King Charles spaniel with a 48-hour history of haematochezia, icterus and collapse. Regenerative anaemia with a packed-cell volume of 7 per cent was seen. Prior to referral, radiography had revealed a gastric, metallic foreign body which was removed at exploratory laparotomy. On presentation, the dog was comatose, hypothermic and bradycardic - resuscitation was performed successfully, but the dog then displayed marked abdominal pain. The dog died 12 hours after presentation. At postmortem examination, the animal showed severe icterus. Both kidneys were diffusely dark red; the pancreas was diffusely pale and nodular. Histopathological examination revealed evidence of intravascular haemolysis with blood vessel lumens containing haemoglobin. The renal tubules also contained large amounts of intraluminal haemoglobin with haemoglobin crystals scattered throughout the cortex and medulla. The pancreas exhibited multifocal coagulative necrosis, surrounded by a neutrophil-dominated inflammatory infiltrate. Zinc levels were markedly increased above the normal reference range in both liver and kidney. This report describes the clinical and pathological findings of a case of acute zinc toxicity in a dog following ingestion of a metallic object which resulted in marked haemolytic anaemia and acute pancreatitis.

A survey of wild-rodent populations has revealed that murine norovirus (MNV) is present and diverse in wild-house mice Mus musculus. This virus is genetically similar to MNV infecting show mice and previously described variants circulating in laboratory mice. The detection of MNV in wild-mouse populations suggests that MNV infection of laboratory mice and show mice (from which laboratory mice are derived) derives from contact with or their origins from wild-mouse progenitors. The survey additionally identified frequent infection of wood mice (Apodemus sylvaticus) with genetically divergent variants of MNV. These viruses are distinct from previously described MNV variants, differing by 22-23 % over the complete genome sequence compared with a maximum of 13 % between M. musculus-derived strains. Comparison with other noroviruses reveals that the Apodemus MNV groups with MNV in genogroup V and shares the same overall genome organization, predicted lengths of proteins encoded by ORFs 1-3 and the existence of a conserved alternative reading frame in VP1 encoding a homologue of the MNV ORF4. Different Apodemus MNV isolates were as variable as MNV isolates and showed evidence for inter-isolate recombination. Our observation of species-specific associations of MNV variants in wild populations suggests that murine noroviruses have an ancient origin, a feature that they may share with other norovirus genogroups. INTRODUCTIONMurine norovirus (MNV) was first identified in laboratory mice deficient in both the recombination-activating gene 2 (RAG2) and the signal transducer and activator of transcription 1 gene (STAT1) (Karst et al., 2003). MNV infection appears to be widespread in laboratory mice, with serological evidence of infection found in 6-26 % of sera (Hsu et al., 2005;Kim et al., 2011) and PCR-positive results obtained in 23 % of samples (6-100 % in different facilities; Kim et al., 2010). MNV has been identified in a wide variety of inbred and transgenic mouse strains from laboratories across the world.The origin of MNV in laboratory mouse strains is unknown. Phylogenetic analysis of the complete genome sequences of 26 MNV variants suggests that MNV comprises a single group (genogroup V) of viruses that are more than 87 % identical to each other (Thackray et al., 2007). However, genetic subgroupings of MNV variants are associated with the animal facility of origin rather than country or mouse strain (Thackray et al., 2007;Kim et al., 2010;Barron et al., 2011). This could reflect sampling of MNV diversity present in the progenitors of modern laboratory mice, or the repeated introduction of MNV virus variants from unknown local hosts (Barron et al., 2011). Serological evidence for MNV infection has been reported for two of 31 wild mice trapped in Pennsylvania, USA (Parker et al., 2009), but no nucleotide sequence information is available to confirm the identity of these isolates.A zoonotic relationship between mice and humans for MNV is unlikely as there are no reports of MNV in humans. Zoonotic relationships for ot...

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