Effect of Preoperative Erythromycin or Dexamethasone/Vitamin C on Postoperative Abomasal Emptying Rate in Dairy Cows Undergoing Surgical Correction of Abomasal Volvulus

Objective— To determine the effects of preoperative erythromycin or combined dexamethasone/vitamin C treatment on postoperative abomasal emptying rate in cows undergoing surgical correction of abomasal volvulus (AV).
Study Design— Prospective, controlled, clinical study using a convenience sample.
Animals— Lactating Holstein–Friesian cows (n=45) with AV were alternately assigned to 3 groups (n=15): group C: untreated (control); group E: erythromycin (10 mg/kg intramuscularly [IM]); group D: dexamethasone (0.02 mg/kg intravenously [IV]) and vitamin C (10 mg/kg IV).
Methods— Drugs were administered 1 hour before surgical correction of AV. d -xylose solution (50%, 0.5 g/kg body weight) was injected into the abomasal lumen during surgery. Jugular venous blood samples for determination of serum d -xylose concentration were periodically obtained. Time to maximal serum d -xylose concentration (T_max-model) was pharmacokinetically determined.
Results— Abomasal emptying rate was significantly ( P <0.05) faster in group E (T_max-model=182±69 min; mean±SD) than in group C cows (T_max-model=237±64 min). Abomasal emptying rate was similar in group D (T_max-model=196±47 min) and group C. Both treatments improved postoperative milk yield within 1 day after surgery.
Conclusion— Preoperative injection of erythromycin (10 mg/kg IM) is an effective method for ameliorating postoperative abomasal hypomotility in cows with AV.
Clinical Relevance— Parenteral erythromycin can be recommended for preoperative treatment of cows with AV.     

Urinary Tract Infection in Dogs with Thoracolumbar Intervertebral Disc Herniation and Urinary Bladder Dysfunction Managed by Manual Expression,Indwelling Catheterization or Intermittent Catheterization

Objective— To evaluate risk factors for lower urinary tract infection (UTI) in dogs with intervertebral disc disease (IVDD) that had manual expression (ME), indwelling catheterization (IDC) or intermittent catheterization (ITC) for urinary bladder management. Study Design— Randomized‐clinical trial. Animals— Dogs (n=62) treated with urinary bladder dysfunction requiring surgery for IVDD and control dogs (n=30) that had surgery for reasons other than IVDD. Methods— Treated dogs were randomly assigned to ME, IDC, or ITC. Urine was collected for culture and antimicrobial susceptibility testing before and after treatment. Incidence and risk factors for UTI were evaluated. Bacterial isolates and antimicrobial resistance patterns were described. Results— Mean (±SD) time to urination was significantly longer for IDC dogs (7.4±2.75 days) than ME dogs (4.2±2.63) and ITC dogs (4.9±3.12). Thirteen treated dogs (21%) and no control dogs developed UTI: 4/25 (16%) ME, 8/25 (32%) IDC, and 1/12 (8%) ITC. Enterobacter sp. was most frequently isolated (4/13; 31%). Duration of treatment was the only risk factor for UTI and each additional day of treatment increased the risk of UTI 1.5 times. Conclusion— For dogs with acute IVDD, the duration of required urinary bladder management establishes the risk of UTI, not the urinary bladder management technique. Clinical Relevance— Duration of treatment for urinary bladder dysfunction is a risk factor for UTI in dogs recovering from acute IVDD. Treatment for urinary bladder management should be limited where possible and no method of treatment is preferred. For dogs managed by IDC, voluntary urination might occur before clinically suspected.     

Aural Cholesteatoma in Twenty Dogs

Objective— To determine the clinical course in dogs with aural cholesteatoma. Study Design— Case series. Animals— Dogs (n=20) with aural cholesteatoma. Methods— Case review (1998–2007). Results— Twenty dogs were identified. Clinical signs other than those of chronic otitis externa included head tilt (6 dogs), unilateral facial palsy (4), pain on opening or inability to open the mouth (4), and ataxia (3). Computed tomography (CT) was performed in 19 dogs, abnormalities included osteoproliferation (13 dogs), lysis of the bulla (12), expansion of the bulla (11), bone lysis in the squamous or petrosal portion of the temporal bone (4) and enlargement of associated lymph nodes (7). Nineteen dogs had total ear canal ablation–lateral bulla osteotomy or ventral bulla osteotomy with the intent to cure; 9 dogs had no further signs of middle ear disease whereas 10 had persistent or recurrent clinical signs. Risk factors for recurrence after surgery were inability to open the mouth or neurologic signs on admission and lysis of any portion of the temporal bone on CT imaging. Dogs admitted with neurologic signs or inability to open the mouth had a median survival of 16 months. Conclusions— Early surgical treatment of aural cholesteatoma may be curative. Recurrence after surgery is associated with advanced disease, typically indicated by inability to open the jaw, neurologic disease, or bone lysis on CT imaging. Clinical Relevance— Presence of aural cholesteatoma may affect the prognosis for successful surgical treatment of middle ear disease.     

Cloning, expression and characterization of monkey (Macaca fascicularis) CD137

CD137 plays an important role as a co-stimulatory molecule in activated T cells. Agonistic CD137 specific antibodies have been investigated as therapeutic agents to promote tumor-specific immune responses by direct activation of T cells. As part of the pre-clinical pharmacological evaluation of cynomolgus monkeys, monkey CD137 was cloned and characterized. The deduced amino acid sequence encoded a full-length gene of 254 amino acids 95% identical to human CD137. Sequence variants identified in monkey CD137 include four splice variants lacking the transmembrane domain. These variants were detectable in human including two previously unreported variants. Two missense single nucleotide polymorphisms were detected present in 42 and 50% of 36 monkeys tested. In both monkey and human, mRNA expression of full-length CD137 and splice variants were significantly increased in peripheral blood mononuclear cells (PBMCs) upon stimulation by anti-CD3 antibodies. Recombinant monkey CD137 protein was bound with high affinity by an agonistic anti-human CD137 antibody but not by an anti-mouse CD137 antibody. In summary, compared to human, monkey CD137 showed distinct extracellular domain amino acid sequence and sequence polymorphisms. Thus, antibodies directed against epitopes in this extracellular domain could have differences in pharmacologic activity between cynomolgus monkeys and human or across individual cynomolgus monkeys.     

Optimization and validation of a flow cytometric method for immunophenotyping peripheral blood lymphocytes from cynomolgus monkeys (Macaca fascicularis)

BACKGROUND: Guidelines published by the Food and Drug Administration and Center for Human Medicinal Products describe the need to assess immunotoxic effects in nonclinical studies that evaluate drug toxicity, including the use of immunophenotyping to measure immunotoxicity. We are not aware of previous studies, however, that have validated methods for immunophenotyping peripheral blood lymphocyte subsets in whole blood samples from cynomolgus monkeys. OBJECTIVE: The purpose of this study was to optimize and validate a flow cytometric assay for immunophenotyping lymphocytes in the peripheral blood of cynomolgus monkeys. METHODS: A series of prevalidation experiments were done to determine optimal reagents, volumes, timing, and other procedural details of the flow cytometric assay. Using the optimized method, we then determined precision, interindividual variation, laboratory-to-laboratory variability, and sample stability. Stabilized human blood was used as a positive control for staining, processing, and analysis. The percentage and number of pan-T cells (CD3+), T-helper cells (CD3+4+), T cytotoxic/suppressor cells (CD3+8+), natural killer cells (CD3-16+), and B-cells (CD3-20+) were determined in 146 male and 140 female, clinically healthy monkeys and reference intervals were calculated. RESULTS: By doing 4-color staining with a lyse-wash method, intra- and interassay precision were <5% for all lymphocyte subsets. Variability between technicians and laboratories was minimal (CVs<3%). Samples were stable for up to 24 hours after staining and fixing. CONCLUSIONS: The validated method is extremely robust and can be performed under good laboratory practice conditions to support nonclinical studies. Reference intervals for lymphocyte subsets were similar to those previously reported.     

Acute myeloid leukemia with multilineage dysplasia in an alpaca

An 18-year-old female alpaca was presented to the Colorado State University Veterinary Teaching Hospital for chronic ill thrift over a 1-year period. Six weeks previously, an infected left mandibular cheek tooth was removed by oral extraction. On physical examination the patient was cachectic, lethargic, and weak. Abnormalities on the CBC included neutropenia, thrombocytosis, and severe nonregenerative, macrocytic, hypochromic anemia. Dysplastic nucleated erythrocytes and micromegakaryocytes were observed on the peripheral blood smear. Neutrophils, bands, and metamyelocytes appeared markedly toxic. Numerous blasts containing variable numbers of fine azurophilic granules were also observed. Based on their morphology, the cells were interpreted to be progranulocytes and myeloblasts, and a presumptive diagnosis of acute myeloid leukemia (AML) was made. The blast cells accounted for 60% of the nucleated cell population on bone marrow aspirates, further supporting a diagnosis of AML with multilineage dysplasia. Post mortem examination showed infiltration of the neoplastic cells into spleen, liver, kidney, and lymph nodes. Based on histologic findings, the morphologic diagnoses were disseminated myeloid neoplasia, chronic regionally extensive tooth root abscess, and membranous glomerulonephritis. The neoplastic cells were CD172a-positive on flow cytometry, chloroacetate esterase-positive by cytochemistry, and myeloperoxidase-positive by immunohistochemistry, confirming myeloid origin. To our knowledge, this is the first case of AML with multilineage dysplasia in an alpaca, with only one other case of myelodysplasia described previously in this species.     

Metastatic immune infiltrates correlate with those of the primary tumour in canine osteosarcoma

Our lack of understanding of the immune microenvironment in canine osteosarcoma (cOSA) has limited the identification of potential immunotherapeutic targets. In particular, our ability to utilize readily available tissue from a dog's primary tumour to predict the type and extent of immune response in their pulmonary metastatic lesions is unknown. We, therefore, collected 21 matched pairs of primary tumours and pulmonary metastatic lesions from dogs with OSA and performed immunohistochemistry to quantify T‐lymphocyte (CD3), FOXP3+ cell, B‐lymphocyte (Pax‐5), and CD204+ macrophage infiltration. We found that T‐lymphocytes and FOXP3+ infiltrates in primary tumours positively correlated with that of metastatic lesions (ρ = 0.512, P = 0.038 and ρ = 0.698, P = 0.007, respectively), while a strong trend existed for CD204+ infiltrates (ρ = 0.404, P = 0.087). We also observed T‐ and B‐lymphocytes, and CD204+ macrophages to be significantly higher in a dog's pulmonary metastasis compared to their primary tumour (P = 0.018, P = 0.018, P = 0.016, respectively), while FOXP3+ cells were only significantly higher in metastases when all primary tumour and metastasis lesions were compared without pairing (P = 0.036). Together, these findings suggest that the metastatic immune microenvironment may be influenced by that of the primary cOSA, and that primary tumour immune biomarkers could potentially be applied to predict immunotherapeutic responses in gross metastatic disease. We, therefore, provide a rationale for the treatment of cOSA pulmonary metastases with immunotherapeutics that enhance the anti‐tumour activity of these immune cells, particularly in dogs with moderate to high immune cell infiltration in their primary tumours.     

Association of macrophage and lymphocyte infiltration with outcome in canine osteosarcoma

Immunotherapeutic strategies have shown promise for the treatment of canine osteosarcoma (cOSA). Very little is known about the immune microenvironment within cOSA, however, limiting our ability to identify potential immune targets and biomarkers of therapeutic response. We therefore prospectively assessed the disease‐free interval (DFI) and overall survival time (ST) of 30 dogs with cOSA treated with amputation and six doses of adjuvant carboplatin. We then quantified lymphocytic (CD3+, FOXP3+) and macrophage (CD204+) infiltrates within the primary tumours of this cohort using immunohistochemistry, and evaluated their association with outcome. Overall, the median DFI and ST were 392 and 455 days, respectively. The median number of CD3+ and FOXP3+ infiltrates were 45.8 cells/mm² (4.6‐607.6 cells/mm²) and 8.5 mm² (0‐163.1 cells/mm²), respectively. The median area of CD204+ macrophages was 4.7% (1.3%‐23.3%), and dogs with tumours containing greater than 4.7% CD204+ macrophages experienced a significantly longer DFI (P = 0.016). Interestingly, a significantly lower percentage of CD204+ macrophages was detected in cOSA arising from the proximal humerus compared to other appendicular bone locations (P = 0.016). Lymphocytic infiltrates did not appear to correlate with outcome in cOSA. Overall, our findings suggest that macrophages may play a role in inhibiting cOSA progression, as has been suggested in human osteosarcoma.