Development and validation of a questionnaire to assess health-related quality-of-life in cats with hyperthyroidism.

BACKGROUND: Health-related quality-of-life (HRQoL) assessment tools are becoming increasingly important for the assessment of diseases in veterinary medicine. OBJECTIVES: To develop a tool to assess the HRQoL of hyperthyroid cats and their owners. ANIMALS: Cats with hyperthyroidism (n = 229) and without hyperthyroidism (n = 322). METHODS: Cross-sectional study design. A preliminary list of 28 questions relating to the HRQoL of hyperthyroid cats and the influence their cat's disease might have on owners was created. Each question consisted of 2 subquestions: (1) "how often does the item apply"; (2) "how strongly does the item affect HRQoL." The questionnaire was refined based on statistical analysis, including Mann-Whitney-U tests on each item, comparing the results from cats with and without hyperthyroidism. Internal consistency and reliability of the questions were measured by Cronbach's alpha (α). P < .05 was considered significant. RESULTS: Overall, 25/28 questions were retained within the final HRQoL tool, which had an excellent internal consistency (α = .92). The tool produced a score between 0 and 382 (lower scores meaning better HRQoL). The median HRQoL score was 87.5 (range, 2-348) for cats with hyperthyroidism, and 27 (range, 0-249) for cats without (P < .001), suggesting the HRQoL was poorer in hyperthyroid cats. CONCLUSIONS AND CLINICAL IMPORTANCE: This validated HRQoL tool is useful to reliably quantify the influence of hyperthyroidism on the quality-of-life of affected cats and their owners. In the future, it could be considered of assistance in the clinical assessment of cats with hyperthyroidism.

Comparison of the Sysmex XT-2000iV with microscopic differential counts of canine bone marrow.

Canine bone marrow is frequently assessed in the advanced preclinical research environment. Automated analysis provides time savings and objectivity over the gold standard of microscopic (cytologic) evaluation. We compared the analysis of 90 canine bone marrow samples by the Sysmex XT-2000iV hematology analyzer (Sysmex Corp., Kobe, Japan) with cytologic evaluation. Gates for cell populations were created in the system's WBC/BASO channel. Variables "total nucleated red blood cells" (total_NRBC), "poly- and orthochromatic nucleated red blood cells" (poly_orth_NRBC), "total neutrophils" (total_NEUT), "mature neutrophils" (mature_NEUT), and myeloid-to-erythroid (M:E) ratio were compared with cytologic evaluation. Intra-assay repeatability and total error (TE) were calculated for both methods. Intra-assay repeatability was 0.95-2.48% for the XT-2000iV and 8.32-23.23% for cytology. Observed TE for the automated measurement was 5.16-46.8% and for cytology 22.70-76.74%. Spearman rank correlation was excellent for M:E ratio (0.91) and fair for the other populations (0.65-0.71). Absolute bias for M:E ratio was low (-0.114). A negative absolute bias of -7.71% for the XT-2000iV was found for poly_orth_NRBC, whereas the bias was positive for total_NEUT (7.10%) and mature_NEUT (14.67%). M:E ratio of canine bone marrow samples can be precisely determined using the Sysmex XT-2000iV WBC/BASO channel. Total_NRBC, poly_orth_NRBC, total_NEUT, and mature_NEUT can be estimated rapidly. With distinctly lower coefficient of variation and observed TE compared with cytology, automated measurement provides advantages in terms of standardization, and it is suited to the advanced preclinical research environment where large numbers of samples are investigated.

Characterization of bone turnover and energy metabolism in a rat model of primary and secondary osteoporosis.

BACKGROUND: An experimental rat model served for evaluation of bone- and energy metabolism in early and late stages of osteoporosis. For the early stage, we hypothesized that bilateral ovariectomy (OVX)+multi-deficiency diet (OVXD; depletion of vitamin D, calcium, vitamin K, phosphorus) would induce increased bone turnover while the late stage would be characterized by enhanced bone catabolism. Obesity, insulin resistance and hyperleptinemia would be seen during the whole course of disease. Healthy female Sprague Dawley rats (n=41) aged 10 weeks were randomly assigned to sham and treatment groups and sacrificed at 3, 12, and 14 months after the study began. RESULTS: In the early phase, OVXD was associated with an increase in body weight, but not, however, in later stages. There was a decrease in bone mineral density and relative bone volume (BV/TV) as assessed by Dual Energy X-ray Absorptiometry and micro computed tomography that was most severe in the later stages of disease, indicating bone catabolism. Osteocalcin limiting bone formation was increased initially, whereas later stages (14 months) were characterized by elevated osteopontin, suggesting bone remodeling. Severe hyperparathyroidism was present during all stages of disease. Only the early phases of disease were characterized by hyperinsulinemia and increased adrenocorticotrophic stimulating hormone, whereas in the late stage hypoleptinemia rather than hyperleptinemia was seen. CONCLUSION: Markers of bone and energy metabolism reflected both an increased bone turn over and ongoing bone remodeling associated with initial hyperinsulinemia. Osteopontin and osteocalcin can be used to differentiate early and late stages of osteoporosis.

Evaluation of the impedance analyzer PocH-100iV Diff for analysis of canine and feline blood.

BACKGROUND: An automated impedance-based in-house hematology analyzer, the PocH-100iV Diff, which provides a 3-part leukocyte differential count that includes eosinophils, recently has been introduced. OBJECTIVES: The aims of this study were to validate results from the PocH-100iV Diff for dogs and cats and evaluate the impact of the anticoagulant used and sample storage conditions. METHODS: Blood samples collected in K(3) EDTA from 153 cats and 150 dogs were included in the comparison study. The reference analyzer was the ADVIA 2120 hematology analyzer, and manual differential leukocyte counts and PCV were the manual reference methods. RESULTS: Coefficients of variation were < 3% except for platelet counts and feline differential and eosinophil counts. Correlation between analyzers was good to excellent except for hemoglobin (HGB) concentration in dogs and RBC indices for both species. Biases were close to 0 except for MCHC and platelet counts. Correlation with manual counts was good for lymphocytes and OTHR cells (combined neutrophil and monocyte counts) and fair and poor for feline and canine eosinophil counts, respectively. Estimated sensitivity and specificity for detection of eosinophilia were, respectively, 50% and 98% for cats and 34% and 77% for dogs. A significant anticoagulant effect was seen for MCV in cats and for HCT, MCH, MCHC, and platelet, OTHR, and eosinophil counts in dogs. RBC and WBC counts, HGB concentration, and MCH were stable for 72 h. HCT, MCV, MCHC, and platelet counts were affected by sample storage (dogs > cats; 22°C > 4°C). CONCLUSIONS: The PocH-100iV Diff is a suitable in-house instrument. A strength is its specific, but moderately sensitive, detection of feline eosinophils.

Effect of submaximal aerobic exercise on platelet function, platelet activation, and secondary and tertiary hemostasis in dogs.

OBJECTIVE: To investigate whether submaximal aerobic exercise in dogs is followed by activation of all phases of coagulation as has been reported for humans. ANIMALS: 9 healthy Beagles. PROCEDURES: 30 minutes before dogs were exercised, a 16-gauge central venous catheter was placed in a jugular vein of each dog by use of the catheter-through-the-needle technique. Samples were collected before exercise, after running on a treadmill (6 km/h for 13 minutes), and at 60 minutes. Platelet activation was evaluated with platelet morphology indices (mean platelet component, mean platelet volume, and number of large platelets) provided by a laser-based hematology system. Platelet function was assessed in hirudin-anticoagulated whole blood with an impedance-based aggregometer with collagen as the agonist (final concentrations, 0, 1.6, 3.2, 5, and 10 µg/mL). Prothrombin time, activated partial thromboplastin time, and concentrations of fibrinogen, factor VIII, antithrombin, protein C, protein S, and fibrin D-dimer were determined automatically. Kaolin-activated thromboelastography variables R (reaction time), K (clot formation time), angle α, maximal amplitude, and G (clot stability) were measured in recalcified citrated whole blood. RESULTS: Exercise resulted in a significant decrease in mean platelet volume and the number of large platelets but did not change the mean platelet component, which reflected platelet activation as well as platelet function. Secondary and tertiary coagulation did not change significantly, nor did thromboelastography variables. CONCLUSIONS AND CLINICAL RELEVANCE: Aerobic exercise resulted in a decrease in the number of large and thus most likely activated platelets but otherwise had no major impact on coagulation in dogs.

Influence of blood collection technique on platelet function and coagulation variables in dogs.

OBJECTIVE: To investigate the influence of diameter of the catheter and blood collection technique on platelet function and variables reflecting secondary hemostasis, physiologic anticoagulation, and fibrinolysis in dogs. ANIMALS: 6 healthy Beagles. PROCEDURES: Blood samples were collected with 20- and 18-gauge venous catheters immediately after catheters were inserted in a peripheral vein, through a 14-gauge central venous catheter that had been placed via the Seldinger technique in a jugular vein < 30 minutes before sample collection, and through a 13-gauge central venous catheter placed via a catheter-through-the-needle technique < 30 minutes before sample collection (techniques 1 to 4, respectively). Platelet function was assessed in hirudin-anticoagulated whole blood via an impedance-based aggregometer with collagen (0.8, 0.4, 0.2, 0.1, and 0.05 µg/mL) as an inductor. Kaolin-activated thromboelastography variables were determined in citrated whole blood. Prothrombin time, activated partial thromboplastin time, fibrinogen and fibrin D-dimer concentrations, and activity of factor VIII, antithrombin, protein C, and protein S were assessed automatically in citrated plasma. RESULTS: At 0.05 µg of collagen/mL, the highest median rate of aggregation was observed for collection techniques 2 and 3 with 4.3 (range, 2.5 to 6.5) and 3.7 (range, 2.8 to 8.3) aggregation units/min; however, these values were not significantly different from values for the other collection techniques. Generally, sample collection technique did not have a significant impact on results of coagulation variables investigated. CONCLUSIONS AND CLINICAL RELEVANCE: Various blood collection techniques can be used to obtain samples for coagulation testing.

Effect of hemolysis on canine kaolin-activated thromboelastography values and ADVIA 2120 platelet activation indices.

BACKGROUND: The impact of hemolysis on thromboelastography (TEG) and platelet activation indices has not been evaluated. OBJECTIVE: The aim of this study was to investigate the influence of hemolysis induced mechanically (HM) and hemolysis induced by freezing (HF) on TEG, platelet counts (PLT), and platelet activation indicators. METHODS: Blood from 17 dogs was divided into the following samples: controls, HM, and HF. HM was induced by 20 repetitions of expulsion of blood through a 23 g needle. Freezing was at -80 degrees C, followed by warming to 37 degrees and dilution with equal parts room temperature blood at 22 degrees C. TEG variables that were examined included reaction time (R), coagulation time (K), angle (alpha), maximum amplitude (MA), and clot rigidity (G). Platelet indices were measured with the ADVIA 2120 hematology analyzer. RESULTS: Hematocrit (HCT) (mean+/-SD) for controls, HM, and HF were 0.41+/-0.02, 0.39+/-0.03, and 0.25+/-0.02 L/L, respectively, consistent with decreases in HCT of 4.8% (HM) and 39.0% (HF). HM resulted in decreased R (2.5+/-0.9 minutes compared with 5.2+/-1.9 minutes for controls; P<0.001), and HF resulted in increased K (15.2+/-8.6 minutes compared with 5.3+/-4.0 minutes in controls; P<0.01) and decreased alpha (20+/-11 degrees compared with 46+/-17 degrees in controls; P<0.001). MA was decreased more in HF samples (26+/-2 mm) than in HM (38+/-8 mm) or control samples (49+/-9 mm; P<0.0001). The same applied to G values. PLT decreased after HM but not after HF. Hemolysis of both types resulted in decreased mean platelet component (MPC) concentration: control, 19.3+/-2.0, HM 15.5+/-3.4, and HF 14.3+/-0.7 g/dL (P<0.0001). CONCLUSION: In hemolyzed samples decreased MPC and R suggested activated primary and secondary hemostasis, respectively, but decreased MA and G indicated reduced clot firmness, possibly due to hyporeactive platelets. TEG and platelet activation indices should be interpreted cautiously after hemolysis.

Biodegradable beta-Tri-Calciumphosphate/hydroxyethyl methacrylate enhanced three component bone adhesive demonstrates biocompatibility without evidence of systemic toxicity in a rabbit model.

Bone gluing is an attractive surgical technique; however, its use in patients is hampered by a variety of side effects. Therefore, it was the aim of this ethically approved study to evaluate a novel biodegradable beta-Tri-Calciumphosphate (beta-TCP, Cerasorb)-enhanced bone adhesive regarding its toxicity and biocompatibility in a rabbit model. Fifty healthy New Zealand White rabbits were assigned in the study (n = 21) and sham-operated control group (n = 29). In the study group, a cylindrical part (4.6 x 10.0 mm) of the proximal tibia and distal femur was removed, reimplanted, and bone adhesive was applied. Blinded physical examination and sampling for hematology, clinical chemistry, and acute phase proteins (haptoglobin, C-reactive protein (CRP)) was performed before surgery and after 12, 24, 48, 72, 240, and 504 h. Significant findings of the physical examination included a slightly higher grading of warmth (p = 0.0019) and pain (p < 0.0001) of the wound 240 h after surgery in the study group. No significant differences between albumin, haptoglobin, CRP, and urea concentrations in both groups were evident, whereas the study group demonstrated significantly lower leukocyte count, total protein, and globulin concentrations (p < 0.0001). As expected, both groups showed a marked transient increase in muscle enzymes (Creatine Kinase and Aspartate Aminotransferase) following the surgery (p < 0.0001). Twelve to 24 h after surgery, a significant decrease in ionized calcium from 1.38 +/- 0.12 mmol/L to 1.06 +/- 0.13 mmol/L was noted in the study group (p < 0.0001). The results clearly indicated that the novel beta-TCP-enhanced bone adhesive showed good biocompatibility without significant evidence of acute or subacute local or systemic toxicity.

Argyrophilic nucleolar organizing regions and Ki67 equally reflect proliferation in fine needle aspirates of normal, hyperplastic, inflamed, and neoplastic canine lymph nodes (n = 101).

BACKGROUND: The count of argyrophilic nucleolar organizing regions (AgNOR) has been considered a useful variable that reflects cellular proliferation in canine lymph nodes, but it has not been compared with other markers of proliferation. HYPOTHESIS: Ki67 and AgNORs are equally useful as markers of tissue proliferation in fine needle aspirates of canine lymph nodes. ANIMALS: A total of 101 dogs. MATERIAL AND METHODS: Prospective, observational study of a convenience sample of dogs. Two smears were prepared for a May-Gruenwald-Giemsa stain and a Ki67/AgNOR double stain. In addition, CD3/CD79a immunostaining was performed when cytologic examination revealed a lymphoma. The dogs were grouped as normal (n = 26), reactive hyperplasia (n = 25), lymphadenitis (n = 31), and lymphoma (n = 19), based on the physical examination and the cytologic findings. The AgNOR count/cell, AgNOR area/cell and the percentage of cells staining positive for Ki67 were evaluated in 100-167 cells (median, 113 cells) by using automatic image analysis. RESULTS: Mean (SD) AgNOR counts/cell were 1.36 +/- 0.19 in normal dogs, 1.55 +/- 0.26 in lymphadenitis, 1.65 +/- 0.32 in reactive hyperplasia, and 3.67 +/- 1.08 in lymphoma. The percentage of Ki67 positive cells was 2.67 +/- 0.99% in normal lymph nodes, 5.04 +/- 3.34% in lymphadenitis, 5.36 +/- 2.14% in reactive hyperplasia, and 30.2 +/- 10.8% in lymphoma. All variables were significantly higher in dogs with lymphoma compared with the other groups (P < .0001). The sensitivity and the specificity of the AgNOR count for diagnosing lymphoma were 95 and 96% at a cutoff value of >2.04 AgNORs/cell. The cutoff value for the Ki67 positive cells was >10.40% (sensitivity, 95%; specificity, 98%). CONCLUSION AND CLINICAL IMPORTANCE: The results indicated that both AgNOR and Ki67 counts were good diagnostic tools for assessment of proliferation in aspirates of canine lymph nodes.

Liver disease in dogs with tracheal collapse.

BACKGROUND: Hepatopathy in dogs with chronic respiratory diseases is poorly recognized. The aim of this study was to evaluate liver parameters alanine transferase, alkaline phosphatase, and glutamate dehydrogenase, as well as basal and stimulated bile acid concentration, in dogs with tracheal collapse. HYPOTHESIS: Dogs with tracheal collapse have hepatopathy. ANIMALS: 26 dogs with tracheal collapse. MATERIALS AND METHODS: Gall bladder contraction was stimulated by intramuscular injection of a synthetic cholecystokinin analogue (ceruletide). Twelve healthy Beagle dogs and 30 dogs of various breeds investigated previously without evidence of hepatic, gastrointestinal, or respiratory diseases served as control. Amelioration of liver variables was assessed after stent implantation. RESULTS: Twelve of 26 (46%) dogs had increased serum activity of 2 or more liver enzymes. Serum basal bile acid concentrations were high in 24 of 26 dogs. Twenty- and 40-minute stimulated bile acids were significantly higher in dogs with tracheal collapse (64.2 +130.0/-43.0 micromol/L and 82.6 +164.0/-57.1 micromol/L) compared to the control dogs (7.0 +/- 3.6 micromol/L and 6.4 +/- 3.5 micromol/L). All twelve dogs reevaluated after a median of 58 days (48-219 days) had a normal breathing pattern and significantly decreased 20 and 40 minutes stimulated bile acids (50.0 +92.7/-32.8 micromol/L, 52.8 +97.6/-34.3 micromol/L; P = .0043), whereas plasma liver enzyme activities were not significantly influenced. CONCLUSION AND CLINICAL IMPORTANCE: There was a significant hepatic dysfunction in the majority of dogs with a tracheal collapse. Liver function should be routinely assessed in dogs with severe respiratory disease.

Cerebrospinal fluid from a 6-year-old dog with severe neck pain.

A 6-year-old, intact female, Labrador Retriever/Terrier cross was presented to the University Veterinary Hospital, University College Dublin with a 3-week history of therapy-resistant cervical pain and intermittent fever. Physical examination findings included marked cervical pain resulting in neck extension and vocalization. Examination of the CSF revealed mild pleocytosis (total nucleated cells = 0.009 x 10(9)/L, reference interval <0.005 x 10(9)/L). Cytocentrifuged preparations of the CSF were of low cellularity, containing predominantly macrophages and occasional small lymphocytes. Several small- to medium-sized fragments of a slightly granular, amorphous, eosinophilic substance were observed. The majority of mononuclear cells were located within this material, in small groups of 3-13 cells. The amorphous foamy material stained positive with Luxol fast blue, suggestive of myelin-like material. The dog was euthanized and postmortem examination revealed intervertebral disk protrusion between C2 and C3. Hematoxylin- and Luxol fast blue-stained histopathologic sections of brain and spinal cord revealed only mild hemorrhage. The extracellular material in the CSF of this dog may have been caused by myelin degeneration or leakage of phospholipids from damaged cells. Because no histologic evidence of demyelination was observed with the disk extrusion, the myelin-like material in this case was thought to be the product of phospholipid breakdown from damaged cellular membranes. Three cases of dogs with spinal cord disease and myelin-like material in the CSF have been reported previously. The clinical significance of this finding is still unknown.

Calcospherite-like bodies and caseous necrosis in tracheal mucus from a dog with tuberculosis.

A 3-year-old Wirehaired Fox Terrier was presented to the University Veterinary Hospital, University College Dublin, for evaluation of chronic cough of 8-months duration. Bronchoscopy showed a severely dilated collapsed left principal bronchus filled with highly viscous white mucus. Cytologically, globular lipid-like material and round concentrically laminated crystalline structures were evident within the proteinaceous mucus. These findings resembled the calcospherites and granular caseous debris often observed in human tuberculous patients. A Ziehl-Neelsen-stained cytocentrifuged preparation of material obtained by bronchoalveolar lavage revealed a few acid-fast rods within macrophages, suggestive of tuberculosis. At necropsy, granulomas with caseous necrosis were present in the lung parenchyma, bronchial and mediastinal lymph nodes, liver, pancreas, and mesentery. Granulomas were adherent to both kidney capsules and to the diaphragm. Histologically, there was evidence of mild calcification within caseous granulomas, which was confirmed by von Kossa's stain. Using Ziehl-Neelsen stain, acid-fast rods were identified within granulomas; bacterial culture was positive for Mycobacterium bovis. The cytologic findings in this case have not been reported previously in dogs and demonstrate a possible correlation between tuberculosis and calcospherite-like bodies with caseous, globular material in bronchial mucus, similar to that described in human patients.