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Abstract
Objective—To establish an in vivo method for matrix metalloproteinase (MMP)-2 and MMP-9 induction in horses via IV administration of lipopolysaccharide (LPS) and to evaluate the ability of doxycycline, oxytetracycline, flunixin meglumine, and pentoxifylline to inhibit equine MMP-2 and MMP-9 production.
Animals—29 adult horses of various ages and breeds and either sex.
Procedures—In part 1, horses received an IV administration of LPS (n = 5) or saline (0.9% NaCl) solution (5). Venous blood samples were collected before and at specified times for 24 hours after infusion. Plasma was harvested and analyzed for MMP-2 and MMP-9 activities via zymography. In part 2, horses received doxycycline (n = 5), oxytetracycline (5), flunixin meglumine (5), or pentoxifylline (4) before and for up to 12 hours after administration of LPS. Plasma was obtained and analyzed, and results were compared with results from the LPS-infused horses of part 1.
Results—Administration of LPS significantly increased MMP-2 and MMP-9 activities in the venous circulation of horses. All MMP inhibitors significantly decreased LPS-induced increases in MMP activities but to differing degrees. Pentoxifylline and oxytetracycline appeared to be the most effective MMP-2 and MMP-9 inhibitors, whereas doxycycline and flunixin meglumine were more effective at inhibiting MMP-2 activity than MMP-9 activity.
Conclusions and Clinical Relevance—IV administration of LPS to horses caused increased venous plasma activities of MMP-2 and MMP-9. These MMP activities were reduced by pentoxifylline and oxytetracycline, suggesting that further evaluation of these medications for treatment and prevention of MMP-associated diseases in horses is indicated.
Abstract
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.
Abstract
Objective—To evaluate the effects of dorsal versus lateral recumbency on the cardiopulmonary system during isoflurane anesthesia in red-tailed hawks (Buteo jamaicensis).
Animals—6 adult 1.1- to 1.6-kg red-tailed hawks.
Procedures—A randomized, crossover study was used to evaluate changes in respiratory rate, tidal volume, minute ventilation, heart rate, mean arterial and indirect blood pressures, and end-tidal Pco 2 measured every 5 minutes plus Paco 2 and Pao 2 and arterial pH measured every 15 minutes throughout a 75-minute study period.
Results—Respiratory rate was higher, tidal volume lower, and minute ventilation not different in lateral versus dorsal recumbency. Position did not affect heart rate, mean arterial blood pressure, or indirect blood pressure, although heart rate decreased during the anesthetic period. Birds hypoventilated in both positions and Paco 2 differed with time and position × time interaction. The Petco 2 position × time interaction was significant and Petco 2 was a mean of 7 Torr higher than Paco 2. The Paco 2 in dorsal recumbency was a mean of 32 Torr higher than in lateral recumbency. Birds in both positions developed respiratory acidosis.
Conclusions and Clinical Relevance—Differences in tidal volume with similar minute ventilation suggested red-tailed hawks in dorsal recumbency might have lower dead space ventilation. Despite similar minute ventilation in both positions, birds in dorsal recumbency hypoventilated more yet maintained higher Pao 2, suggesting parabronchial ventilatory or pulmonary blood flow distribution changes with position. The results refute the hypothesis that dorsal recumbency compromises ventilation and O2 transport more than lateral recumbency in red-tailed hawks.
Abstract
Objective—To evaluate vascular endothelial growth factor (VEGF) concentrations in canine blood products treated with or without a leukoreduction filter.
Sample—10 canine blood donors.
Procedures—Dogs underwent blood collection. Five of 10 units were leukoreduced prior to separation into packed RBCs and fresh frozen plasma (FFP). Concentrations of VEGF were measured by ELISA in plasma supernatants from aliquots of packed RBCs obtained immediately after separation and on days 7, 14, and 21 of storage. Fresh frozen plasma samples of 2 filtered and 2 nonfiltered units were examined after storage.
Results—RBC counts in whole blood before and after leukoreduction did not differ significantly, but WBCs and platelets were removed effectively. The VEGF concentration was lower than the detection limit (9 pg/mL) in 9 of 10 plasma samples and in all packed RBC and FFP units immediately after separation. The median VEGF concentrations in 5 nonfiltered packed RBC units were 37, 164, and 110 pg/mL on days 7, 14, and 21 of storage, respectively. In 5 filtered packed RBC and all FFP units, VEGF concentrations remained lower than the detection limit.
Conclusions and Clinical Relevance—Leukoreduction filters were effective in preventing the release of VEGF during storage of canine RBC products.
Abstract
Objective—To investigate the effects of high-intensity training (HIT) on carbohydrate and fat metabolism in Thoroughbreds.
Animals—12 Thoroughbreds (3 to 4 years old; 6 males and 6 females).
Procedures—Horses performed HIT for 18 weeks. They ran at 90% or 110% of maximal oxygen consumption (
o
2max) for 3 minutes (5 d/wk) and were subjected to incremental exercise testing (IET) before and after training. Blood samples were collected during IET, and muscle samples were obtained from the gluteus medius muscle immediately after IET. Phosphofructokinase, citrate synthase, and β-3-hydroxyacyl CoA dehydrogenase (β-HAD) activities were measured to determine glycolytic and oxidative capacities. Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and fatty acid translocase (FAT/CD36) protein contents were detected via western blotting. Metabolome analysis was performed via capillary electrophoresis–electrospray ionization mass spectrometry to measure substrate concentrations related to carbohydrate metabolism.
Results—Peak speed during IET and
o
2max increased after HIT. Activities of citrate synthase and β-HAD increased after HIT, whereas phosphofructokinase activity remained unchanged. The PGC-1α and FAT/CD36 protein contents increased after HIT, but plasma lactate concentration and the respiratory exchange ratio decreased after HIT. The plasma free fatty acid concentration increased after HIT, whereas the glucose concentration was not altered. Fructose 1,6-diphosphate, phosphoenolpyruvate, and pyruvate concentrations decreased after HIT.
Conclusions and Clinical Relevance—HIT caused an increase in oxidative capacity in equine muscle, which suggested that there was a decreased reliance on carbohydrate utilization and a concomitant shift toward fatty acid utilization during intensive exercise.
Abstract
Objective—To compare values of lower urogenital tract urodynamic and morphometric variables determined during the prepubertal (sexually immature) period and first and second estrous cycles in healthy female Beagle littermates to determine functional and anatomic changes of the lower urogenital tract during those periods.
Animals—5 female Beagle littermates.
Procedures—Urethral pressure profilometry, diuresis cystometry, and vaginourethrography were performed when dogs were 3.5, 4.5, 5, 6, 7, 8, 8.5, and 9 months old and during proestrus; estrus; early, middle, and late diestrus; and early and late anestrus of the first and second estrous cycles.
Results—At the end of the prepubertal period, values of urodynamic and morphometric variables increased significantly, compared with values at earlier times. Maximum bladder capacity developed when dogs were 9 months old. In all dogs, the bladder was intermittently located in an intrapelvic position during the prepubertal period; the bladder was intra-abdominal from the time dogs were 9 months old until the end of the study. Urethral pressure decreased significantly during estrus and early diestrus of the first and second estrous cycles. Bladder capacity increased significantly during diestrus of both estrous cycles. Urethral and vaginal lengths were significantly longer during proestrus and estrus than they were during anestrus.
Conclusions and Clinical Relevance—Values of lower urogenital tract urodynamic and morphometric variables were influenced by age and phases of the estrous cycle of immature and young adult Beagles in this study. Age of dog and phase of estrous cycle should be considered when interpreting urodynamic and vaginourethrography data.
Abstract
Objective—To examine the effect of endotoxins on metabolism and histopathologic changes of isolated perfused equine forelimbs.
Sample—Forelimbs (comprising the metacarpus and digit) were collected from cadavers of 12 healthy adult horses after slaughter at an abattoir (14 limbs; 1 forelimb of 10 horses and both forelimbs of 2 horses).
Procedures—Forelimbs were perfused for 10 hours with autologous blood, with and without the addition of endotoxin (80 ng of lipopolysaccharide [LPS]/L). Two limbs of the endotoxin exposure group and 2 nonperfused limbs were loaded to failure of the suspensory apparatus of the pedal bone to evaluate the effect of body weight. Metabolic and histologic variables were evaluated.
Results—Blood pressure increased during the first hour and did not differ between groups. Lactate dehydrogenase activity was similar in both groups and increased significantly during the 10-hour period; glucose consumption at 5 hours and lactate concentration at 8 hours were significantly higher in limbs exposed to endotoxin. The width of secondary epidermal lamellae was greater in LPS limbs. In the primary dermal lamellae of LPS limbs, there were significantly more vessels with an open lumen and aggregates of intravascular neutrophils.
Conclusions and Clinical Relevance—In the blood-perfused isolated forelimbs of equine cadavers, exposure to LPS led to significant changes in the laminar tissue as well as to metabolic changes. Therefore, endotoxin should be considered as a causative factor for laminitis and not merely as a risk factor.
Abstract
Objective—To determine associations between serum concentrations of omega-3 polyunsaturated fatty acids or body condition and serum concentrations of adiponectin, leptin, insulin, glucose, or triglyceride in healthy dogs.
Animals—62 healthy adult client-owned dogs.
Procedures—Body condition score and percentage of body fat were determined. Blood samples were collected after food was withheld for 12 hours. Serum was harvested for total lipid determination, fatty acid analysis, and measurement of serum concentrations of adiponectin, leptin, insulin, glucose, and triglyceride. Associations between the outcome variables (adiponectin, leptin, insulin, glucose, and triglyceride concentrations) and each of several variables (age, sex, percentage of body fat, and concentrations of total lipid, α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid) were determined.
Results—Serum concentrations of docosapentaenoic acid were significantly positively associated with concentrations of adiponectin and leptin and negatively associated with concentrations of triglyceride. Serum concentrations of α-linolenic acid were significantly positively associated with concentrations of triglyceride. No significant associations were detected between serum concentrations of eicosapentaenoic acid or docosahexaenoic acid and any of the outcome variables. Percentage of body fat was significantly positively associated with concentrations of leptin, insulin, and triglyceride but was not significantly associated with adiponectin concentration. Age was positively associated with concentrations of leptin, insulin, and triglyceride and negatively associated with concentrations of adiponectin. Sex did not significantly affect serum concentrations for any of the outcome variables.
Conclusions and Clinical Relevance—Docosapentaenoic acid may increase serum concentrations of adiponectin and leptin and decrease serum triglyceride concentration in healthy dogs.
Abstract
Objective—To evaluate effects of extracellular lactate on viability, shape change, lactate metabolism, and reactive oxygen species (ROS) production in equine polymorphonuclear leukocytes (PMNs).
Sample—PMNs isolated from equine venous blood samples.
Procedures—PMNs were incubated with 0 to 300mM lactate for 30 minutes before each experiment. Viability was assessed via trypan blue exclusion. Shape change was assessed via flow cytometry and light microscopy. Relative quantification of monocarboxylic acid transporter and lactate dehydrogenase lactate dehydrogenase (LDH) isotype mRNAs was performed with a real-time PCR assay. Effects of lactate at a pH of 7.4 to 6.0 on ROS production in response to phorbol 12-myristate 13-acetate, opsonized zymosan, or N-formyl-methionyl-leucyl-phenylalanine was assessed by luminol-dependent chemiluminescence.
Results—Lactate had no effect on viability of PMNs but did alter their size and density. Monocarboxylic acid transporter 1 and lactate dehydrogenase B mRNA values were not altered. Monocarboxylic acid transporter 4 and lactate dehydrogenase A mRNA values were significantly decreased. Lactate incubation of cells significantly decreased PMN-derived luminol-dependent chemiluminescence and induced different sensitivities to stimulants (phorbol 12-myristate 13-acetate, opsonized zymosan, and N-formyl-methionyl-leucyl-phenylalanine). The response ratio to N-formyl-methionyl-leucyl-phenylalanine revealed that PMNs were primed by incubation with up to 50mM lactate, significantly increasing the production of ROS. Incubation with lactate and acidic pH caused a synergistic effect on ROS production.
Conclusions and Clinical Relevance—Extracellular lactate potentially has a direct effect on the capacity to produce ROS by equine PMNs, which may be associated with alterations in innate immune functions within a short period after high-intensity exercise.
Abstract
Objective—To establish a safe anesthetic protocol with little effect on blood biochemical values and IV glucose tolerance test (IVGTT) results in Japanese black bears (Ursus thibetanus japonicus).
Animals—16 captive female Japanese black bears (5 to 17 years of age).
Procedures—Bears were randomly assigned to 4 treatment groups (4 bears/group) in which various treatment combinations were administered via blow dart: tiletamine HCl and zolazepam HCl (9 mg/kg) alone (TZ), TZ (6 mg/kg) and acepromazine maleate (0.1 mg/kg), TZ (6 mg/kg) and butorphanol tartrate (0.3 mg/kg), or TZ (3 mg/kg) and medetomidine HCl (40 μg/kg). Glucose injection for the IVGTT was started 130 minutes after TZ administration. Blood samples were obtained before, at, and intermittently after glucose injection for measurement of biochemical variables as well as plasma glucose and serum insulin concentrations during the IVGTT. Rectal temperature, pulse rate, and respiratory rate were assessed every 15 minutes during the experiment.
Results—Induction and maintenance of anesthesia were safely achieved with little adverse effect on cardiopulmonary function when each of the 4 anesthetic regimens was used, although mild hypothermia was induced. No difference was evident between treatment groups in blood biochemical values. Blood glucose and insulin concentration profiles during the IVGTT were similar among the bears given TZ, with or without acepromazine or butorphanol, but hyperglycemia and hypoinsulinemia developed in bears given TZ with medetomidine.
Conclusions and Clinical Relevance—All 4 anesthetic regimens yielded chemical restraint without affecting clinical and biochemical values in bears, but medetomidine appeared to affect IVGTT results. For this reason, medetomidine should not be used when anesthetizing bears for IVGTTs.
