Sixty cases of canine sialocoele were reviewed from the records of the Sydney University Veterinary Teaching Hospital. Poodles, dachshunds and Australian silky terriers were significantly over represented in the survey when compared to the hospital sample population (P&0.01). No other statistically significant breed, sex, or age trends were noted. Neoplasia was associated with sialocoele formation in three cases. The sialocoele location was cervical (42), sublingual (four), pharyngeal (four), not recorded (one) or in more than one site (nine). Thirty‐five of 39 surgically treated cases were managed by extirpation of the mandibular and sublingual salivary glands together with drainage. Few complications were recorded. Forty‐two per cent of the cases had received previous veterinary treatment in the form of drainage alone. Drainage of saliva from sialocoeles without removal of the offending salivary glands is inappropriate.

No abstract

Death Associated Protein kinase (DAP kinase) a novel calmodulin-dependent serine/threonine kinase was ®rst identi®ed as a positive mediator of programmed cell death. Loss of DAP kinase expression was ®rst demonstrated in highly metastatic cells, whilst reexpression of the protein resulted in delayed local tumour growth and a decreased incidence of metastasis. Although loss of DAP kinase expression has been reported in several cell lines derived from human malignancies the mechanisms responsible have not been de®ned. In this study we have examined 32 sporadic pituitary tumours for expression of the DAP kinase protein and transcript. In addition, we examined the methylation and deletion status of the DAP kinase CpG island as possible mechanisms for the inactivation of the DAP kinase gene. Eleven of 32 (34%) tumours had undetectable DAP kinase expression, by Western blot and/or RT ± PCR analysis. Loss of DAP kinase expression was signi®cantly (P=0.004) associated with invasive tumours (10 of 17; 59%) compared to their noninvasive (1 of 15; 7%) counterparts. Of 11 tumours that failed to express DAP kinase, ®ve (45%) showed de novo methylation of the CpG island contained within the promoter region, while four (36%) had evidence of homozygous deletion of this region. Statistical analysis showed that loss of DAP kinase expression was signi®cantly (P=50.001) associated with methylation or deletion of the DAP kinase CpG island. With two exceptions, none of the remaining tumours or ®ve histologically normal post-mortem pituitaries examined had evidence of methylation or deletion within this region. To our knowledge this is the ®rst report that describes two mutually exclusive mechanisms associated with loss of DAP kinase gene expression. In addition, we also show that loss of the DAP kinase protein and associated genetic aberrations preferentially segregates with tumours that show an invasive phenotype.

BackgroundThe hCMV promoter is very commonly used for high level expression of transgenes in mammalian cells, but its utility is hindered by transcriptional silencing. Large genomic fragments incorporating the CpG island region of the HNRPA2B1 locus are resistant to transcriptional silencing.Results In this report we describe studies on the use of a novel series of vectors combining the HNRPA2B1 CpG island with the hCMV promoter for expression of transgenes in CHO-K1 cells. We show that the CpG island gives at least twenty-fold increases in the levels of EGFP and EPO observed in pools of transfectants, and that transgene expression levels remain high in such pools for more than 100 generations. These novel vectors also allow facile isolation of clonal CHO-K1 cell lines showing stable, high-level transgene expression.ConclusionVectors incorporating the hnRPA2B1 CpG island give major benefits in transgene expression from the hCMV promoter, including substantial improvements in the level and stability of expression. The utility of these vectors for the improved production of recombinant proteins in CHO cells has been demonstrated.

Summary A retrospective study of 119 cases of fracture of the proximal phalanx is described. The short incomplete sagittal fracture (split pastern) was most common and carried a good prognosis for a return to racing following conservative treatment. Longer incomplete fractures also had a good prognosis but complete fractures, either through the lateral cortex of the proximal phalanx or through the bone into the proximal interphalangeal joint, required internal fixation. Careful assessment of these and comminuted fractures was essential before surgery was undertaken. Most comminuted fractures were treated with a view to the animal retiring to stud rather than racing again. Humane destruction was considered to be justifiable for such fractures in colts or geldings of no stud value or if the fracture was severely comminuted.

Some chlorine-resistant Escherichia coli isolates harbor the locus of heat resistance (LHR), a genomic island conferring heat resistance. In this study, the protective effect of the LHR for cells challenged by chlorine and oxidative stress was quantified. Cloning of the LHR protected against NaClO (32 mM; 5 min), H2O2 (120 mM; 5 min), and peroxyacetic acid (105 mg/liter; 5 min) but not against 5.8 mM KIO4, 10 mM acrolein, or 75 mg/liter allyl isothiocyanate. The lethality of oxidizing treatments for LHR-negative strains of E. coli was about 2 log10 CFU/ml higher than that for LHR-positive strains of E. coli. The oxidation of cytoplasmic proteins and membrane lipids was quantified with the fusion probe roGFP2-Orp1 and the fluorescent probe BODIPY581/591, respectively. The fragment of the LHR coding for heat shock proteins protected cytoplasmic proteins but not membrane lipids against oxidation. The middle fragment of the LHR protected against the oxidation of membrane lipids but not of cytoplasmic proteins. The addition of H2O2, NaClO, and peroxyacetic acid also induced green fluorescent protein (GFP) expression in the oxidation-sensitive reporter strain E. coli O104:H4 Δstx2::gfp::amp. Cloning of pLHR reduced phage induction in E. coli O104:H4 Δstx2::gfp::amp after treatment with oxidizing chemicals. Screening of 160 strains of Shiga toxin-producing E. coli (STEC) revealed that none of them harbors the LHR, additionally suggesting that the LHR and Stx prophages are mutually exclusive. Taking our findings together, the contribution of the LHR to resistance to chlorine and oxidative stress is based on the protection of multiple cellular targets by different proteins encoded by the genetic island. IMPORTANCE Chlorine treatments are used in water and wastewater sanitation; the resistance of Escherichia coli to chlorine is thus of concern to public health. We show that a genetic island termed the locus of heat resistance (LHR) protects E. coli not only against heat but also against chlorine and other oxidizing chemicals, adding to our knowledge of the tools used by E. coli to resist stress. Specific detection of the oxidation of different cellular targets in combination with the cloning of fragments of the LHR provided insight into mechanisms of protection and demonstrated that different fragments of the LHR protect different cellular targets. In E. coli, the presence of the LHR virtually always excluded other virulence factors. It is tempting to speculate that the LHR is maintained by strains of E. coli with an environmental lifestyle but is excluded by pathogenic strains that adapted to interact with vertebrate hosts.

Measuring the activity and temperature of rats is commonly required in biomedical research. Conventional approaches necessitate single housing, which affects their behavior and wellbeing. We have used a subcutaneous radiofrequency identification (RFID) transponder to measure ambulatory activity and temperature of individual rats when group-housed in conventional, rack-mounted home cages. The transponder location and temperature is detected by a matrix of antennae in a baseplate under the cage. An infrared high-definition camera acquires side-view video of the cage and also enables automated detection of vertical activity. Validation studies showed that baseplate-derived ambulatory activity correlated well with manual tracking and with side-view whole-cage video pixel movement. This technology enables individual behavioral and temperature data to be acquired continuously from group-housed rats in their familiar, home cage environment. We demonstrate its ability to reliably detect naturally occurring behavioral effects, extending beyond the capabilities of routine observational tests and conventional monitoring equipment. It has numerous potential applications including safety pharmacology, toxicology, circadian biology, disease models and drug discovery.

The technique described in this report permits simple ligation of a range of different ductus morphologies in dogs of varying breeds, weights, and ages. In the event of serious hemorrhage, prompt ventricular outflow occlusion and ductus ligation followed by rapid whole blood transfusion is life saving in most cases.