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Flow cytometric detection and procoagulant activity of circulating canine platelet-derived microparticles
2013
Helmond, Sarah E. | Catalfamo, James L. | Brooks, Marjory B.
Objective: To measure platelet membrane–derived microparticle (PMP) content and thrombin-generating capacity of canine plasma subjected to specific processing and storage conditions. Animals: 31 clinically normal dogs (19 males and 12 females). Procedures: Citrate-anticoagulated blood samples obtained from each dog were centrifuged at 2,500 × g to isolate platelet-poor plasma (PPP), then PPP was centrifuged at 21,000 × g to isolate microparticle-free plasma (MPF) and microparticle-enriched plasma (MPEP). Whole blood and paired samples of fresh and frozen-thawed PPP, MPF, and MPEP were dual labeled for flow cytometric detection of membrane CD61 (constitutive platelet antigen) and annexin V (indicating phosphatidylserine externalization). Platelets and PMPs were enumerated with fluorescent, size-calibrated beads. Thrombin generation in fresh and frozen-thawed PPP, MPF, and MPEP was measured via kinetic fluorometric assays configured with low tissue factor and low phospholipid concentrations. Results: Initial centrifugation yielded PPP with < 0.5% the platelets of whole blood, with median counts of 413 PMPs/μL for males and 711 PMPs/μL for females. Sequential centrifugation resulted in a 10-fold concentration of PMPs in MPEP and virtually depleted PMPs from MPF. Thrombin generation depended on PMP content, with median endogenous thrombin potential of 0, 893, and 3,650 nmol•min for MPF, PPP, and MPEP, respectively. Freeze-thaw cycling caused significant increases in PMP counts and phosphatidylserine externalization. Conclusions and Clinical Relevance: Canine PMPs were major determinants of thrombin-generating capacity; preanalytic variables influenced plasma PMP content. Processing conditions described here may provide a basis for characterization of PMPs in clinical studies of thrombosis in dogs.
Show more [+] Less [-]Evaluation of tissue Doppler imaging for regional quantification of radial left ventricular wall motion in healthy horses
2013
Decloedt, Annelies | Verheyen, Tinne | Sys, Stanislas | Clercq, Dominique de | Loon, Gunther van
Objective: To compare the feasibility and repeatability of tissue Doppler imaging (TDI) for quantification of radial left ventricular (LV) velocity and deformation from different imaging planes and to correlate cardiac event timing data obtained by TDI to M-mode and pulsed-wave Doppler-derived time intervals in horses. Animals: 10 healthy adult horses. Procedures: Repeated echocardiography was performed by 2 observers from right and left parasternal short-axis views at papillary muscle and chordal levels. The TDI measurements of systolic and diastolic velocity, strain rate, strain peak values, and timing were performed in 8 LV wall segments (LV free wall and interventricular septum from right parasternal views; left and right region of LV wall from left parasternal views). The inter- and intraobserver within- and between-day variability and measurement variability were assessed. The correlation between TDI-based measurements and M-mode and pulsed-wave Doppler-based time measurements was calculated. Results: TDI measurements of velocity, strain rate, and strain were feasible in each horse, although deformation could often not be measured in the LV free wall. Systolic and diastolic time intervals could be determined with low to moderate variability, whereas peak amplitude variability ranged from low to high. The TDI-based time measurements were significantly correlated to M-mode and pulsed-wave Doppler measurements. Conclusions and Clinical Relevance: TDI measurements of radial LV velocity and deformation were feasible with low to moderate variability in 8 LV segments. These measurements can be used for evaluating LV function in further clinical studies.
Show more [+] Less [-]Power of treatment success definitions when the Canine Brief Pain Inventory is used to evaluate carprofen treatment for the control of pain and inflammation in dogs with osteoarthritis
2013
Brown, Dorothy Cimino | Bell, Margie | Rhodes, Linda
Objective—To determine the optimal method for use of the Canine Brief Pain Inventory (CBPI) to quantitate responses of dogs with osteoarthritis to treatment with carprofen or placebo. Animals—150 dogs with osteoarthritis. Procedures—Data were analyzed from 2 studies with identical protocols in which owner-completed CBPIs were used. Treatment for each dog was classified as a success or failure by comparing the pain severity score (PSS) and pain interference score (PIS) on day 0 (baseline) with those on day 14. Treatment success or failure was defined on the basis of various combinations of reduction in the 2 scores when inclusion criteria were set as a PSS and PIS ≥ 1, 2, or 3 at baseline. Statistical analyses were performed to select the definition of treatment success that had the greatest statistical power to detect differences between carprofen and placebo treatments. Results—Defining treatment success as a reduction of ≥ 1 in PSS and ≥ 2 in PIS in each dog had consistently robust power. Power was 62.8% in the population that included only dogs with baseline scores ≥ 2 and 64.7% in the population that included only dogs with baseline scores ≥ 3. Conclusions and Clinical Relevance—The CBPI had robust statistical power to evaluate the treatment effect of carprofen in dogs with osteoarthritis when protocol success criteria were predefined as a reduction ≥ 1 in PIS and ≥ 2 in PSS. Results indicated the CBPI can be used as an outcome measure in clinical trials to evaluate new pain treatments when it is desirable to evaluate success in individual dogs rather than overall mean or median scores in a test population.
Show more [+] Less [-]Load redistribution in walking and trotting Beagles with induced forelimb lameness
2013
Abdelhadi, Jalal | Wefstaedt, Patrick | Galindo-Zamora, Vladimir | Anders, Alexandra | Nolte, Ingo | Schilling, Nadja
Objective: To evaluate the load redistribution mechanisms in walking and trotting dogs with induced forelimb lameness. Animals: 7 healthy adult Beagles. Procedures: Dogs walked and trotted on an instrumented treadmill to determine control values for peak and mean vertical force as well as verticle impulse for all 4 limbs. A small sphere was attached to the ventral pad of the right forelimb paw to induce a reversible lameness, and recordings were repeated for both gaits. Additionally, footfall patterns were assessed to test for changes in temporal gait variables. Results: During walking and trotting, peak and mean vertical force as well as vertical impulse were decreased in the ipsilateral forelimb, increased in the contralateral hind limb, and remained unchanged in the ipsilateral hind limb after lameness was induced. All 3 variables were increased in the contralateral forelimb during trotting, whereas only mean vertical force and vertical impulse were increased during walking. Stance phase duration increased in the contralateral forelimb and hind limb during walking but not during trotting. Conclusions and Clinical Relevance: Analysis of the results suggested that compensatory load redistribution mechanisms in dogs depend on the gait. All 4 limbs should be evaluated in basic research and clinical studies to determine the effects of lameness on the entire body. Further studies are necessary to elucidate specific mechanisms for unloading of the affected limb and to determine the long-term effects of load changes in animals with chronic lameness.
Show more [+] Less [-]In vitro effects of lipid emulsion on platelet function and thromboelastography in canine blood samples
2013
Tonkin, Laura R. | Parnell, Nolie K. | Hogan, Daniel F.
Objective-To determine whether soybean oil emulsion has an in vitro effect on platelet aggregation and thromboelastography in blood samples obtained from healthy dogs. Animals-12 healthy adult dogs. Procedures-Blood samples were collected from each dog into tubes containing EDTA, hirudin, or sodium citrate for a CBC, collagen- and ADP-induced impedance aggregometry, or thromboelastography, respectively. Whole blood platelet aggregation, determined with ADP or collagen agonists, was measured in blood samples containing hirudin and final lipid concentrations of 0, 1, 10, and 30 mg/mL. The thromboelastographic variables R (reaction time), K (clotting time), α angle, and maximum amplitude were evaluated in blood samples containing sodium citrate and final lipid concentrations equivalent to those used for assessment of platelet aggregation. Results-Median maximum ADP- and collagen-induced platelet aggregation in blood samples containing 1, 10, or 30 mg of lipid/mL did not differ significantly from the value for the respective lipid-free blood sample. Maximum amplitude determined via thromboelastography was significantly reduced in blood samples containing 10 and 30 mg of lipid/mL, compared with findings for lipid-free blood samples. Values of other thromboelastographic variables did not differ, regardless of lipid concentrations. Conclusions and Clinical Relevance-Maximum amplitude determined via thromboelastography in canine blood samples was significantly affected by the addition of lipid to final concentrations that are several orders of magnitude higher than clinically relevant lipid concentrations in dogs. Lipid treatment appears to have no significant effect on hemostatic variables in dogs, although clinical studies should be performed to confirm these in vitro findings.
Show more [+] Less [-]Antinociceptive effects of long-acting nalbuphine decanoate after intramuscular administration to Hispaniolan Amazon parrots (Amazona ventralis)
2013
Sanchez-Migallon Guzman, David | Braun, Jana M. | Steagall, Paulo V.M. | Keuler, Nicholas S. | Heath, Timothy D. | Krugner-Higby, Lisa A. | Brown, Carolyn S. | Paul-Murphy, Joanne R.
Objective: To evaluate the thermal antinociceptive effects and duration of action of nalbuphine decanoate after IM administration to Hispaniolan Amazon parrots (Amazona ventralis). Animals: 10 healthy adult Hispaniolan Amazon parrots of unknown sex. Procedures: Nalbuphine decanoate (33.7 mg/kg) or saline (0.9% NaCl) solution was administered IM in a randomized complete crossover experimental design (periods 1 and 2). Foot withdrawal threshold to a noxious thermal stimulus was used to evaluate responses. Baseline thermal withdrawal threshold was recorded 1 hour before drug or saline solution administration, and thermal foot withdrawal threshold measurements were repeated 1, 2, 3, 6, 12, 24, 48, and 72 hours after drug administration. Results: Nalbuphine decanoate administered IM at a dose of 33.7 mg/kg significantly increased thermal foot withdrawal threshold, compared with results after administration of saline solution during period 2, and also caused a significant change in withdrawal threshold for up to 12 hours, compared with baseline values. Conclusions and Clinical Relevance: Nalbuphine decanoate increased the foot withdrawal threshold to a noxious thermal stimulus in Hispaniolan Amazon parrots for up to 12 hours and provided a longer duration of action than has been reported for other nalbuphine formulations. Further studies with other types of nociceptive stimulation, dosages, and dosing intervals as well as clinical trials are needed to fully evaluate the analgesic effects of nalbuphine decanoate in psittacine birds.
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