Effect of inclusion or exclusion of epithelial cells in equine respiratory cytology analysis.

Published studies vary as to whether epithelial cells are included in differential counts for tracheal wash (TW) and bronchoalveolar lavage (BAL) cytology in horses. The aim of this study was to determine whether inclusion or exclusion of epithelial cells affects interpretation of airway cytology. Using criteria of >20% TW neutrophils, >10% BAL neutrophils and/or >5% BAL mast cells to indicate airway inflammation, there was a change in categorisation from 'normal' to 'abnormal' in 21%, 4% and 8% horses, respectively, when epithelial cells were excluded from differential counts. It is recommended that future equine respiratory research studies explicitly state whether epithelial cells are included or excluded in differential counts. A consensus on epithelial cell inclusion during cytology reporting is required.

Basal serum cortisol concentration as a screening test for hypoadrenocorticism in dogs.

BACKGROUND: Measurement of basal serum or plasma cortisol concentration is used as a screening test for hypoadrenocorticism in dogs, but is not well characterized. OBJECTIVES: To evaluate the sensitivity and specificity of basal serum cortisol to detect hypoadrenocorticism in a population of dogs with a clinical suspicion of hypoadrenocorticism. ANIMALS: Four hundred and fifty dogs with nonadrenal gland illness and 14 dogs with naturally occurring hypoadrenocorticism were included. METHODS: Retrospective case-control study. The records of all dogs having had an ACTH stimulation test performed between January 2005 and September 2011 at the University of Bristol were reviewed. Dogs were included if the test was performed as a screening for hypoadrenocorticism. The sensitivity and specificity of basal serum cortisol concentration to detect dogs with hypoadrenocorticism were calculated using 2 cut-offs and compared to the gold standard ACTH stimulation test. RESULTS: Using a cut-off of ≤2 µg/dL (≤55 nmol/L), the sensitivity and specificity of basal cortisol to detect hypoadrenocorticism were 100% and 63.3%, respectively, whereas for a cut-off of ≤1 µg/dL (≤28 nmol/L), the sensitivity and specificity were 85.7% and 91.8%, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Measurement of basal serum cortisol is useful as a screening test for hypoadrenocorticism in dogs using a cut-off of ≤2 µg/dL (≤55 nmol/L), and the disease is unlikely with a basal serum cortisol >2 µg/dL (>55 nmol/L). A basal serum cortisol ≤2 µg/dL (≤55 nmol/L) cannot be used to diagnose hypoadrenocorticism, and an ACTH stimulation test should be performed in these cases.

Comparison of Measurements of 12 Analytes in Equine Blood Samples Using the In-Practice Falcor 350 and the Reference KoneLab 30i Analysers.

Falcor 350 is a wet-reagent biochemistry analyser that is available for in-house use. The aim of this study was to compare the results produced by this analyser with those obtained by the KoneLab 30i that served as the reference instrument. Blood samples from 60 clinical cases were analysed for urea, creatinine, total proteins, albumin, creatine kinase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, total calcium, phosphate, sodium, and potassium using both instruments. Good to excellent correlations (r s value) value) were identified for creatinine (0.88), total proteins (0.92), albumin (0.93), creatine kinase (0.98), aspartate aminotransferase (0.98), alkaline phosphatase (0.94), total bilirubin (0.98), phosphate (0.95), and potassium (0.97). The correlations for total calcium (0.71), sodium (0.68), and urea (0.64) were fair. For albumin, aspartate aminotransferase, creatine kinase, phosphate, potassium, total bilirubin, creatinine, and total proteins, the two instruments produce values that are closely related to each other and are sufficiently similar to allow them to be used interchangeably without the need for additional correction factor computations. Because of differences in the methodologies, the Falcor results for alkaline phosphatase, total calcium, and sodium cannot be used interchangeably and should be interpreted using reference intervals established from the Falcor analyser.

Real-time quantitative polymerase chain reaction detection of haemoplasmas in healthy and unhealthy dogs from Central Macedonia, Greece.

OBJECTIVES: The aims of this study were to determine the prevalence of canine haemoplasmas, Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum" infection in Central Macedonia, Greece, and to evaluate any associations between canine haemoplasma infection and clinical presentation, selected laboratory data or the presence of ticks. METHODS: Genomic DNA was purified from excess blood (n=151) submitted for haematological examination. Purified DNA was subjected to species-specific quantitative polymerase chain reaction assays duplexed with a canine DNA control quantitative polymerase chain reaction. Clinical records were retrospectively examined and selected clinical parameters were compared to haemoplasma infection status. RESULTS: Nine samples were excluded due to inadequate canine DNA polymerase chain reaction results. Of the remaining 142 samples: eight (5·6%) were positive for M. haemocanis alone, six (4·2%) were positive for "Ca. M. haematoparvum" alone and one (0·7%) was dual positive. No association was found between haemoplasma status and age, sex, breed, health status, presence of anaemia, selected biochemistry parameters, presence of ectoparasites, routine ectoparasiticide treatment or the presence of selected tick-borne diseases.

Effect of storage time on automated cell count and cytological interpretation of body cavity effusions.

The effect of 24- and 48-hour storage at room temperature on automated total nucleated cell count (TNCC), differential cell count (DCC) and cell morphology was assessed, and the effect of initial total protein concentration on canine and feline body cavity effusion samples (2 to 5 ml) was evaluated. At 24 and 48 hours, TNCC and absolute numbers of neutrophils, macrophages and small lymphocytes were significantly decreased and numbers of unrecognisable cells were significantly increased. Neoplastic cells and intracellular bacteria identified in fresh samples were missed at 24 and 48 hours. The initial total protein concentration was associated with an effect on percentage of unrecognisable cells and small lymphocytes over time. Change in TNCC over time would have resulted in misclassification of the effusion type in four of 47 samples.

Septic pericardial effusion associated with pulmonary and pericardial botryomycosis in a dog.

The authors report a case of septic pericardial effusion resulting in cardiac tamponade associated with intrathoracic botryomycosis in a dog. Septic pericarditis and a pulmonary mass were diagnosed, and subtotal pericardiectomy and lobectomy of the affected pulmonary areas were carried out. Histopathology of the excised tissue showed changes supportive of botryomycosis--namely a pyogranulomatous inflammation with neutrophils centred around amorphous homogeneous eosinophilic material and club-like bodies containing Gram-positive bacterial cocci present in the centre. The patient recovered well following surgery and antibiotic therapy. To the authors' knowledge, this is the first report of pulmonary botryomycosis in the dog and the first report of this condition presented with pericardial involvement and cardiac tamponade in any species.

Activity assays for characterizing the thrombolytic protein fibrolase.

Characterization of the thrombolytic agent fibrolase was accomplished employing specific proteolytic and thrombolytic assays. This paper describes a method to measure enzyme proteolytic activity using the oxidized beta-chain of insulin as a substrate. Advantages of this method include a short incubation time for substrate cleavage followed by an isocratic HPLC method with a retention time of approx. 5 min. Proteolytic activity can be rapidly and easily quantitated with this procedure. An azocasein assay was also used to quantitate proteolytic activity. This method was optimized with respect to substrate concentration and incubation time allowing for the rapid quantitation of fibrolase activity. A thrombolytic assay is described which employs fibrin plate clearance and has the advantage of rapid and accurate quantitation compared with previously described methods. It also allows for the standardization of fibrolase in plasmin-equivalent units.

Laugh it off. The effect of humor on the well-being of the older adult.

1. Laughter releases excessive physical and psychological energy, and it reduces stress, anxiety, worry, and frustration. Humor as a skillful nursing intervention can enhance the overall well-being of older adults. 2. If the timing is inappropriate, humor can be a destructive rather than constructive intervention, especially in nursing situations where embarrassing, sensitive, or emotion-laden issues are at hand. 3. This study found that as one ages, one's self-perception of health seems to decrease. Therefore, nurses need to promote a positive outlook toward the aging process and one's own health status while interacting with the elderly in a variety of settings. 4. Nurses should be encouraged to explore the use of humor, not only with patients of all ages, but also with their nurse colleagues to help reduce burn out; to cope with the everyday stress and pressures inherent in the profession; and to enhance learning in the classroom.

Biochemical characteristics of fibrolase, a fibrinolytic protease from snake venom.

Fibrolase, a fibrinolytic enzyme isolated from Agkistrodon c. contortrix (southern copperhead) venom, solubilizes fibrin primarily by rapid hydrolysis of the alpha and beta chains. Fibrolase is also an A alpha, B beta fibrinogenase. The breakdown products of fibrin and fibrinogen following incubation with fibrolase were different from those observed with plasmin. This enzyme is a metalloprotease that was inhibited by ethylenediaminetetraacetic acid. Fibrolase was inhibited by dithiothreitol, suggesting that disulfide bonds are important for catalytic activity. It was also inhibited by alpha 2-macroglobulin, but not by the soybean or lima bean trypsin inhibitors, diisopropylfluorophosphate, or p-hydroxymercuribenzoate. Unlike thrombolytic agents such as streptokinase, fibrolase does not activate plasminogen as evidenced by the use of plasmin-specific chromogenic substrate S-2251 and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Biological and thrombolytic properties of fibrolase--a new fibrinolytic protease from snake venom.

Fibrolase, a direct-acting fibrinolytic enzyme has been shown to cleave primarily the A alpha and B beta chains of human fibrin. We have previously reported that fibrolase also exhibits fibrinogenolytic activity and acts mainly as an alpha-chain fibrinogenase. In contrast to the action of streptokinase (plasminogen activator), fibrolase does not activate plasminogen. In vitro thrombolytic efficacy of fibrolase was determined by monitoring the release of radiolabel from iodinated fibrin and human blood clots. Fibrolase effectively digested the clots in a dose-dependent manner. The in vivo efficacy of fibrolase was evaluated in an animal model of arterial thrombosis. Fibrolase was found to be efficacious at dissolving femoral arterial clots following a single intravenous bolus administration. Time to reperfusion was dose dependent and similar to that observed with streptokinase. No adverse effects on blood pressure and heart rate were observed.