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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.
Afficher plus [+] Moins [-]Effects of pneumoperitoneum induced at various pressures on cardiorespiratory function and working space during laparoscopy in cats
2013
Mayhew, Philipp D. | Pascoe, Peter J. | Kass, Philip H. | Shilo-Benjamini, Yael
Objective—To evaluate the effect of pneumoperitoneum on cardiorespiratory variables and working space during experimental induction of 3 intra-abdominal pressures (IAPs) in cats. Animals—6 healthy young adult neutered male domestic shorthair cats. Procedures—All cats were anesthetized through use of a standardized protocol. A catheter was placed in the right femoral artery for blood pressure and blood gas monitoring. A thermodilution catheter was placed in the right jugular vein via fluoroscopic guidance. Cardiopulmonary variables were measured before (baseline) and 2 and 30 minutes after initiation of pneumoperitoneum at IAPs of 4, 8, and 15 mm Hg; these were created through the use of a mechanical insufflator. At each IAP, abdominal dimensions (height, width, and circumference) were measured at a standardized location. Results—At 4 mm Hg and 8 mm Hg IAP, no clinically important changes were identified in cardiorespiratory values. Heart rate, cardiac index, and stroke volume index remained unchanged throughout the study at all IAPs. Mean arterial blood pressure began to increase at 8 mm Hg and was significantly higher, compared with baseline, at both time points at 15 mm Hg. At 15 mm Hg, Paco2 was significantly higher and cats were more acidotic than at baseline. Working space was subjectively greater at 8 mm Hg than at 4 mm Hg IAP; however, at 15 mm Hg, no clinically important enlargement of the working space was identified, compared with at 8 mm Hg. Conclusions and Clinical Relevance—Values of cardiopulmonary variables were largely unchanged by induction of pneumoperitoneum in healthy cats up to an IAP of 8 mm Hg, and no clinically important increases in working space were evident at an IAP of 15 versus 8 mm Hg. These findings provide little justification for use of IAPs > 8 mm Hg in healthy cats undergoing laparoscopic procedures; however, whether the situation is similar in diseased or elderly cats remains to be determined.
Afficher plus [+] Moins [-]Pharmacokinetics of intravenously and orally administered meloxicam in sheep
2013
Stock, Matthew L. | Coetzee, Johann F. | KuKanich, Butch | Smith, Billy I.
Objective: To determine the pharmacokinetics of meloxicam after IV and PO administration to 6 healthy sheep. Animals: 6 healthy adult Dorset cross sheep (5 males and 1 female). Procedures: Meloxicam (0.5 mg/kg, IV, or 1.0 mg/kg, PO) was administered in a randomized crossover design with a 10-day washout period. Blood samples were collected at predetermined times over 96 hours. Serum drug concentrations were determined by high-pressure liquid chromatography with mass spectrometry. Computer software was used to estimate values of pharmacokinetic parameters through noncompartmental methods. Results: Following IV administration (n = 5), the geometric mean (range) elimination half-life was 14.0 hours (10.5 to 17.0 hours), volume of distribution was 0.204 L/kg (0.171 to 0.272 L/kg), and clearance was 0.17 mL/min/kg (0.12 to 0.27 mL/min/kg). Following oral administration (n = 6), maximum serum concentration was 1.72 μg/mL (1.45 to 1.93 μg/mL), time to maximum serum concentration was 19.0 hours (12.0 to 24.0 hours), clearance per bioavailability was 0.22 mL/min/kg (0.16 to 0.30 mL/min/kg), and terminal half-life was 15.4 hours (13.2 to 17.7 hours). Bioavailability of orally administered meloxicam was calculated as 72% (40% to 125%; n = 5). No adverse effects were evident following meloxicam administration via either route. Conclusions and Clinical Relevance: Meloxicam administered PO at 1.0 mg/kg has good bioavailability with slow elimination kinetics in sheep. These data suggested that meloxicam may be clinically useful, provided the safety and analgesic efficacy of meloxicam as well as feed-related influences on its pharmacokinetics are established in ruminants.
Afficher plus [+] Moins [-]Effects of interstitial cystitis on the acoustic startle reflex in cats
2013
Hague, Devon W. | Stella, Judi L. | Buffington, C.A Tony
Objective: To compare acoustic startle reflexes (ASRs) of healthy cats and cats with interstitial cystitis (IC). Animals: 28 healthy cats (11 males and 17 females) and 20 cats with IC (13 males and 7 females). Procedures: To evaluate the effect of neutering on ASRs, ASRs in neutered and unneutered healthy cats were measured. To evaluate the effect of housing facility acclimation on ASRs in cats with IC, ASRs were measured in cats with IC within 1 month after arrival at the housing facility and again 2 to 3 months after arrival. To evaluate the effect of the environment on ASRs, ASRs were evaluated in all cats with and without IC after acclimation but before and then after environmental enrichment. Results: Neutering led to a significant decrease in overall ASR in the healthy cats. Habituation to the housing facility resulted in a significant decrease in overall ASR of female but not male cats with IC. Environmental enrichment led to a significant decrease in ASR in cats with IC but not in healthy cats. Conclusions and Clinical Relevance: The magnitude of the ASR appeared to be sensitive to environmental conditions and affected by sex, both in healthy cats and cats with IC. It was also higher in cats with IC versus healthy cats, except when cats were housed in a highly enriched environment. Impact for Human Medicine: Treatment approaches that include reduction of a patient's perception of environmental unpredictability may benefit humans with IC.
Afficher plus [+] Moins [-]Influence of acidifying or alkalinizing diets on bone mineral density and urine relative supersaturation with calcium oxalate and struvite in healthy cats
2013
Bartges, Joseph W. | Kirk, Claudia A. | Cox, Sherry K. | Moyers, Tamberlyn D.
Objective—To evaluate the influence of acidifying or alkalinizing diets on bone mineral density and urine relative supersaturation (URSS) with calcium oxalate and struvite in healthy cats. Animals—6 castrated male and 6 spayed female cats. Procedures—3 groups of 4 cats each were fed diets for 12 months that differed only in acidifying or alkalinizing properties (alkalinizing, neutral, and acidifying). Body composition was estimated by use of dual energy x-ray absorptiometry, and 48-hour urine samples were collected for URSS determination. Results—Urine pH differed significantly among diet groups, with the lowest urine pH values in the acidifying diet group and the highest values in the alkalinizing diet group. Differences were not observed in other variables except urinary ammonia excretion, which was significantly higher in the neutral diet group. Calcium oxalate URSS was highest in the acidifying diet group and lowest in the alkalinizing diet group; struvite URSS was not different among groups. Diet was not significantly associated with bone mineral content or density. Conclusions and Clinical Relevance—Urinary undersaturation with calcium oxalate was achieved by inducing alkaluria. Feeding an alkalinizing diet was not associated with URSS with struvite. Bone mineral density and calcium content were not adversely affected by diet; therefore, release of calcium from bone caused by feeding an acidifying diet may not occur in healthy cats.
Afficher plus [+] Moins [-]Effect of acute exercise on monocarboxylate transporters 1 and 4 in untrained and trained Thoroughbreds
2013
Kitaoka, Yu | Endo, Yukari | Mukai, Kazutaka | Aida, Hiroko | Hiraga, Atsushi | Takemasa, Tohru | Hatta, Hideo
Objective-To evaluate the effects of a single incremental exercise test (IET) on mRNA expression and protein content of monocarboxylate transporter (MCT) 1 and MCT4 in the gluteus medius muscle of Thoroughbreds. Animals-12 Thoroughbreds (6 males and 6 females; age, 3 to 4 years). Procedures-Horses underwent an IET before and after 18 weeks of high-intensity exercise training (HIT). Horses were exercised at 90% of maximal oxygen consumption for 3 minutes during the initial 10 weeks of HIT and 110% of maximal oxygen consumption for 3 minutes during the last 8 weeks of HIT. Gluteus medius muscle biopsy specimens were obtained from horses before (baseline), immediately after, and at 3, 6, and 24 hours after the IET. Results-Expression of MCT1 and MCT4 mRNA was upregulated at 3 and 6 hours after the IET in muscle specimens obtained from horses prior to HIT (untrained horses) and at 6 hours after the IET in muscle specimens obtained from horses after HIT (trained horses). For both untrained and trained horses, MCT1 and MCT4 protein contents were increased at 6 hours after the IET and did not differ at 24 hours after the IET, compared with those at baseline. Conclusions and Clinical Relevance-Results indicated that a single IET resulted in transient increases in MCT1 and MCT4 mRNA expression and protein content in untrained and trained horses. These results may be important for the elucidation of exercise-induced alterations in lactate metabolism.
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