Results from the treatment of advanced stage squamous cell carcinoma of the nasal planum in cats, using a combination of intralesional carboplatin and superficial radiotherapy: a pilot study

Six cats with an advanced stage squamous cell carcinoma (SCC) of the nasal planum were treated with a combination of superficial radiotherapy and intralesional carboplatin therapy. This multimodality protocol was well tolerated by the majority of cats and resulted in complete responses in all cats (100%). Median follow‐up for all cats is 268 days, and the median time‐to‐recurrence, time‐to‐progression and overall survival have not yet been reached. Our study, although limited in number of animals and with a relatively short median follow‐up compared to other studies for this disease, suggests that a combination of radiotherapy and intralesional carboplatin is a useful treatment option for an advanced stage SCC of the nasal planum in cats and warrants further application of the multimodality approach presented here.     

RNA‐transfected CD40‐activated B cells as a vaccine strategy in canine malignancies

Introduction: Cell‐based vaccine strategies using dendritic cells as cellular adjuvant have entered phase III trials in humans and have been found to be safe, feasible, and potentially efficacious. Canine patients are generally smaller than adult human patients, which makes production of canine dendritic cell (DC) vaccines problematic, given patient size and the small number of available DC precursors. Here we describe feasibility studies of a novel cell‐based vaccine strategy which uses CD40‐activated B‐cells (CD40‐B) loaded with RNA. This strategy is based on our observations that RNA‐transfected human CD40‐B can drive anti‐tumor T cell responses. One advantage of using CD40‐B cells is the ability to expand this cell population ex vivo, allowing for the numbers of cells required for therapeutic vaccines. Methods: Twenty milliliters of blood were drawn from 6 normal dogs and 5 canine lymphoma patients. Peripheral blood mononuclear cells were separated by Ficoll centrifugation. Culture conditions for B cell activation were optimized using CD40‐ligand, canine IL‐4, and Toll‐like receptor stimulus with CpGoligodinucleotides (ODN). Cyclosporine was added to eliminate peripheral T lymphocytes. Proliferation and activation of CD40‐B cells were demonstrated by CFSE dilution of B cells quantified by flow cytometry. Gene transfer was achieved by mRNA electroporation. Results: Marked in vitro stimulation and proliferation of canine peripheral B cells were achieved with soluble trimeric CD40L, canine IL‐4, and ODN. CD40‐B cells showed dramatic upregulation of MHC class II molecules and CD21 (B‐cell activation marker). After two weeks in culture, cells were negative for CD3 and CD4. Canine CD40‐B cells were efficiently transfected with mRNA, with >60% of CD40‐B expressing green fluorescent protein after GFP mRNA electroporation. Conclusion: RNA‐transfected CD40‐B cells can be efficiently generated from normal and tumor‐bearing dogs. These results provide rationale to test tumor RNA‐transfected CD40‐B as a novel therapeutic approach to treating canine malignancies. Clinical trials in canine lymphoma have been proposed.     

Association of markers in the bovine CAPN1 gene with meat tenderness in large crossbred populations that sample influential industry sires

Two previously identified single-nucleotide polymorphism markers located within the micromolar calcium-activated neutral protease gene (CAPN1) were evaluated for their association with variation in meat tenderness using one commercial sample of Simmental x Angus crossbred calves and one multibreed, crossbred research herd. The commercial sample included 362 animals sired by 23 registered Simmental bulls bred to unregistered Angus cows and represented current industry animals in which to test the predictive merit of the markers. The second sample was a research herd including 564 steers from the Germplasm Evaluation Cycle VII population at the U.S. Meat Animal Research Center, produced with semen from popular sires of the seven Bos taurus beef breeds with the most registrations in the United States (Angus, Charolais, Gelbvieh, Hereford, Limousin, Red Angus, and Simmental) on Angus, Hereford, and MARC III cows. These animals form a relatively outbred population that constituted a stringent test of the predictive merit of the genetic markers, although small groups were half-sibs. Warner-Bratzler shear force measurements were used to determine tenderness phenotypes for all animals. The populations were genotyped for two markers that predict variation at amino acid positions 316 and 530 of the mu-calpain polypeptide, produced by the CAPN1 gene. Minor allele frequencies for markers 316 and 530 in the commercial sample were 0.17 and 0.37, respectively, and in the Cycle VII animals, were 0.20 and 0.28, respectively. Both markers showed association with shear force in the commercial sample (P = 0.04) and the Cycle VII population (P = 0.02), supporting the hypothesis that they represent potential markers to aid selection for improved meat tenderness in commercial populations of beef cattle in the United States.     

The use of KIT and tryptase expression patterns as prognostic tools for canine cutaneous mast cell tumors

Cutaneous mast cell tumors (MCTs) are one of the most common tumors in dogs. Currently, prognostic and therapeutic determinations for MCTs are primarily based on the histologic grade of the tumor, but a vast majority of MCTs are of an intermediate grade, and the prognostic relevance is highly questioned. A more detailed prognostic evaluation, especially of grade 2 canine MCTs, is greatly needed. To evaluate the prognostic significance of KIT and tryptase expression patterns in canine cutaneous MCTs, we studied 100 cutaneous MCTs from 100 dogs that had been treated with surgery only. The total survival and disease-free survival time and the time to local or distant recurrence of MCTs were recorded for all dogs. Using immunohistochemistry, 98 of these MCTs were stained with anti-KIT and antitryptase antibodies. Three KIT- and three tryptase-staining patterns were identified. The KIT-staining patterns were identified as 1) membrane-associated staining, 2) focal to stippled cytoplasmic staining with decreased membrane-associated staining, and 3) diffuse cytoplasmic staining. The tryptase-staining patterns were identified as 1) diffuse cytoplasmic staining, 2) stippled cytoplasmic staining, and 3) little to no cytoplasmic staining. Based on univariate and multivariate survival analysis, increased cytoplasmic KIT staining was significantly associated with an increased rate of local recurrence and a decreased survival rate. The tryptase-staining patterns were not significantly associated with any survival parameter. On the basis of these results, we propose a new prognostic classification of canine cutaneous MCTs, according to their KIT-staining pattern, that can be used for the routine prognostic evaluation of canine cutaneous MCTs.     

Resection cystoplasty of a squamous cell carcinoma in a mare

A 22‐year‐old Arabian mare was examined for evaluation of haematuria of 2 months' duration. Complete blood cell count and chemistry revealed anaemia of chronic disease (i.e. normocytic, normochromic) and hypoproteinaemia. Cystoscopy revealed a haemorrhagic mass protruding into the lumen of the urinary bladder in the apical region. The mass was surgically removed by partial cystectomy performed through a caudal ventral midline laparotomy. Histological examination of the mass confirmed the diagnosis of squamous cell carcinoma. Tumours of the equine urinary bladder are rare, and when a partial cystectomy is indicated, a caudal ventral midline laparotomy may provide adequate surgical exposure even in adult horses.     

A single screw technique compared to a two screw and wire technique as a temporary transphyseal bridge for correction of fetlock varus deformities

A single screw technique is described as a temporary transphyseal bridge for the treatment of fetlock varus angular limb deformity in foals. This has been compared to tension band wiring with regards to rate of correction, cosmetic result and incidence of complications. The single screw was found to be an effective technique resulting in more rapid improvement of deviations in foals aged up to 5 months. Complications were minimal following both techniques and the final cosmetic result was better following single screw surgery.     

Effect of floor cooling and dietary amino acids content on performance and behaviour of lactating primiparous sows during summer

Fifty nine primiparous sows PIC Camborough 23 were distributed in a completely randomized 2 × 2 (with and without floor cooling × two dietary treatments) factorial design with 16 sows/treatment, each sow being considered as an experimental unit. Four replicates of sixteen sows each were used during the trial with the objective of evaluating the effects of floor cooling and the use of dietary amino acid contents on their performance and behaviour during summer. The sows were distributed among the treatments according to body weight and backfat thickness after farrowing. The sows were maintained in the experiment until weaning at 21 days of lactation. The two experimental diets supplied the same levels of crude protein (22%), metabolizable energy (ME; 14.65 MJ/kg) and levels of essential digestible AA relative to digestive lysine and differed according to the digestible lysine to ME ratio (0.75 vs. 0.82 g/MJ of ME). The temperature of the water circulating in the cooled floor was maintained at about 17 °C. Based on the average minimum and maximum temperatures (21.5 and 29.5 °C) obtained during the experimental trial, it can be assumed that the sows were exposed to periods of heat stress. The replicate and the interaction between replicate and treatment effects on all the measurements were not significant. Similarly, no effect of diet or interaction between diet and floor cooling system was found for all criteria measured. An effect (P < 0.05) of floor cooling on average daily feed intake was observed and floor cooling sows showed a higher average (P < 0.05) digestible lysine (61.5 vs. 51.8 g/d) and ME (78.2 vs. 65.9 MJ/d) intakes. The sows submitted to floor cooling showed, consistently, higher absolute values for average weight (+ 8.5 kg) and backfat (+ 0.75 mm) at weaning, compared with the control sows. The sows submitted to the cooled floor showed a shorter (P < 0.01) weaning-to-oestrus interval. The piglet and litter's daily weight gain (DWG), average weight at weaning (AWW) and total weight gain during lactation (TWG) were higher (P < 0.01) for the floor cooling sows. The floor cooling sows showed a higher (P < 0.01) daily milk production. The respiratory rate and rectal temperature values were lower (P < 0.01) for the floor cooling sows. There were differences (P < 0.01) on the cutaneous temperatures measured on the different parts of the sow's body, with the animals submitted to the cooled floor having lower values. The sows submitted to floor cooling spent less (P < 0.01) time in lateral recumbency inactive, more time nursing (P < 0.05) and more time feeding (P < 0.01) compared with control sows. The floor cooling under the sows increased daily feed intake and lysine intake, leading to a lower body weight loss, a lower weaning-to-oestrus interval and also improved nursing behaviour of the sows, leading to a higher milk production and, consequently, higher weight gains of piglets and litter during the lactation period.     

Investigation of the prevalence of neurologic equine herpes virus type 1 (EHV-1) in a 23-year retrospective analysis (1984–2007)

A single nucleotide polymorphism in the equine herpesvirus 1 (EHV-1) DNA polymerase gene (ORF30 A₂₂₅₄ to G) has been associated with clinical signs of equine herpes myeloencephalopathy (EHM). The purpose of our study was to determine the odds ratio for this genetic marker and EHM using a panel of field isolates from North America collected over the past twenty-three years. EHV-1 isolates cultured at the Cornell University Animal Health Diagnostic Laboratory from 1984 to 2007 were retrieved along with their clinical histories. DNA was extracted from these EHV-1 cultures and allelic discrimination was performed using real-time PCR. The results were confirmed by sequencing of the target region in ORF30. PCR and sequencing were in 100% agreement and showed that 19 out of the 176 isolates had the ORF30 G₂₂₅₄ allele (11%), of which16 were EHM cases and 3 respiratory or abortion cases. The odds of having neurologic disease with the ORF30 G₂₂₅₄ genotype were computed as 162 times greater than those with the opposite allele ORF30 A₂₂₅₄ (95% confidence interval: 35–742). Despite this strong statistical significance, 24% (5/21) of horses with neurologic disease in our study population harbored the “non-neurologic” form of the allele (ORF30 A₂₂₅₄), suggesting that other factors may also contribute to the onset of EHM.     

Assessment of the effects of adjunctive gabapentin on postoperative pain after intervertebral disc surgery in dogs

ObjectiveTo assess the effect of adjunctive gabapentin (GBP) on pain after thoracolumbar intervertebral disc surgery in dogs.Study designProspective, randomized, controlled, clinical, ‘blinded’ trial.AnimalsSixty-three client owned dogs undergoing hemilaminectomyMethodsDogs were assigned to two treatment groups. The GBP group received gabapentin 10 mg kg^?1 orally every 12 hours starting before anaesthesia; the placebo (P) group received empty gelatin capsules. Background analgesia was initiated with intravenous levomethadone 0.6 mg kg^?1 (as the combination ‘L-Polamivet) at anaesthesia induction, followed by a fentanyl patch and levomethadone 0.2 mg kg^?1 subcutaneously every 8 hours for 24 hours. Pain was assessed by the short form of the Glasgow Composite Measure Pain Score (CMPS-SF) without the gait category, and by a Visual Analogue Scale (VAS). Serum GBP concentrations and cortisol concentrations were measured. Statistical analyses utilized chi square test, Kolmogorov-Smirnov test, two-way analysis of variances for repeated measurements, Wilcoxon test and Friedmann test as relevant. Correlations were tested by Spearman's and Pearson's correlation coefficient. p< 0.05 was considered significant.ResultsMedian CMPS-SF was lower in group GBP than in group P on days 0.5, 1, 4 and 5. However, CMPS-SF and VAS were not significantly different between groups. Both pain scores decreased significantly over time. Cortisol concentrations were not significantly different between groups. Minimum serum concentrations of GBP fell below the detection limit of 1 μg mL^?1 in 6 of 29 and 7 of 28 dogs at 24 and 72 hours, respectively.Conclusions and clinical relevance10 mg kg^?1 GBP orally twice a day did not result in a detectable reduction in pain behaviour compared to background opioid analgesia alone, although a trend to lower pain levels (p < 0.1) was present. Further studies are needed to determine if this is related to effective background analgesia or an ineffective dose of GBP.