Objective—To evaluate the effects of clopidogrel on clinical and clinicopathologic variables in healthy horses with experimentally induced endotoxemia. Animals—12 adult mares. Procedures—Horses were assigned with a randomization procedure to receive clopidogrel (4 mg/kg, once, then 2 mg/kg, q 24 h; n = 6) or a placebo (6) through a nasogastric tube. After 72 hours of treatment, horses received lipopolysaccharide (LPS; 30 ng/kg, IV). Heart rate, respiratory rate, rectal temperature, CBC variables, plasma fibrinogen concentration, serum tumor necrosis factor-α concentration, plasma von Willebrand factor concentration, and measures of platelet activation (including ADP- and collagen-induced platelet aggregation and closure times, thrombelastography variables, and results of flow cytometric detection of platelet membrane P-selectin, phosphatidylserine, and microparticles) were determined at various times before and after LPS administration by investigators unaware of the treatment groups. Statistical analyses were performed with repeated-measures ANOVA. Results—4 of 6 clopidogrel-treated horses had significant decreases in ADP-induced platelet aggregation before and after LPS administration. Heart rate increased significantly after LPS administration only for the placebo group. No significant differences were detected between groups for CBC variables, closure time, and plasma concentration of fibrinogen or serum concentration of tumor necrosis factor-α, and no clinically relevant differences were detected for other hemostatic variables. Conclusions and Clinical Relevance—In this study, administration of LPS did not induce platelet hyperreactivity in horses on the basis of measures of platelet adhesion, aggregation, degranulation, and procoagulant activity. Administration of clopidogrel was associated with variable platelet antiaggregatory activity and attenuated some clinical signs of endotoxemia.

Objective—To develop a flow cytometric assay to quantify platelet-derived microparticles (PMPs) in equine whole blood and plasma. Sample—Citrate-anticoagulated whole blood from 30 healthy adult horses. Procedures—Platelet-poor plasma (PPP) was prepared from fresh whole blood by sequential low-speed centrifugation (twice at 2,500 × g). Samples of fresh whole blood and PPP were removed and stored at 4° and 24°C for 24 hours. Platelet-derived microparticles were characterized in fresh and stored samples on the basis of the forward scatter threshold (log forward scatter < 10(1)) and labeling with annexin V (indicating externalized phosphatidylserine) and CD61 (a constitutive platelet receptor). A fluorescent bead–calibrated flow cytometric assay was used to determine microparticle counts. Platelet counts, prothrombin time, and activated partial thromboplastin time were measured in fresh samples. Results—Significantly more PMPs were detected in fresh whole blood (median, 3,062 PMPs/μL; range, 954 to 13,531 PMPs/μL) than in fresh PPP (median, 247 PMPs/μL; range, 104 to 918 PMPs/μL). Storage at either temperature had no significant effect on PMP counts for whole blood or PPP. No significant correlation was observed between PMP counts and platelet counts in fresh whole blood or PPP or between PMP counts and clotting times in fresh PPP. Conclusions and Clinical Relevance—Results indicated that the described PMP protocol can be readily used to quantify PMPs in equine blood and plasma via flow cytometry. Quantification can be performed in fresh PPP or whole blood or samples stored refrigerated or at room temperature for 24 hours.

The unfolded protein response (UPR), a conserved cellular response to stressors such as hypoxia and nutrient deprivation, is associated with angiogenesis and metastasis in tumor cells. This article discusses a pilot study conducted to determine whether components of the UPR could be identified in spontaneous canine tumors and whether they were up-regulated within tumor tissue compared with adjacent normal tissue. Tissue samples of various spontaneous canine neoplasms were taken from 13 dogs shortly after surgical excision or euthanasia; control samples were taken from adjacent normal tissue. RNA purification and real-time quantitative reverse-transcription polymerase chain reaction were done to measure the expression of 4 genes associated with the UPR (HERP, CHOP, GRP78, and XBP1s). The results indicated that UPR gene expression can be identified in spontaneous canine tumors and that the UPR is up-regulated, as indicated by significantly increased expression of CHOP and GRP78 within the tumor.

The purpose of the study was to compare the conductance and mannitol permeability of canine colonic mucosa in response to carprofen or 2,4-dinitrophenol (DNP) with or without tempol pretreatment. Ten colonic mucosa sections per dog were mounted in Ussing chambers. Treatments were done in duplicate. Mucosa was exposed to carprofen (200 μg/mL) or DNP (0.25 mM), both with and without tempol (1 mM) pretreatment. Conductance was calculated every 15 min for 240 min. Mannitol flux was calculated over 3 consecutive 60-minute periods. Histology or electron microscopy was done after exposure. Conductance over time, mannitol flux, frequency of histologic categories, and electron microscopic changes were analyzed for treatment effects. The mean ± standard deviation (SD) conductance over time for carprofen or DNP-treated colons was not significantly different from control regardless of tempol pretreatment. Period 3 mannitol fluxes for carprofen and DNP-treated colon were not significantly different, but were greater than control. Period 3 mannitol flux for tempol + carprofen was significantly less than tempol + DNP-treated colon. Sloughing of cells and erosions were seen in the mucosa of carprofen-treated colon. Mitochondrial damage was seen more often in carprofen-treated than DNP-treated or control colon. Tempol pretreatment resulted in more ruptured mitochondria in the carprofen-treated colon; however, other mitochondrial changes were not significantly affected by tempol pretreatment in either carprofen or DNP treated colon. Treatment with carprofen or DNP increased the mannitol flux, but pretreatment with tempol mitigated the carprofen effect. It is apparent that structural mitochondrial damage occurs in the canine colonic mucosa after carprofen and DNP exposure.

Objective - To compare accuracy of a noninvasive single-plane fluoroscopic technique with radiostereometric analysis (RSA) for determining 3-D femorotibial poses in a canine cadaver with normal stifle joints. Sample- Right pelvic limb from a 25-kg adult mixed-breed dog. Procedures- A CT scan of the limb was obtained before and after metal beads were implanted into the right femur and tibia. Orthogonal fluoroscopic images of the right stifle joint were acquired to simulate a biplanar fluoroscopic acquisition setup. Images were obtained at 5 flexion angles from 110° to 150° to simulate a gait cycle; 5 cycles were completed. Joint poses were calculated from the biplanar images by use of RSA with CT-derived beaded bone models and compared with measurements obtained by use of CT-derived nonbeaded bone models matched to single-plane, lateral-view fluoroscopic images. Single-plane measurements were performed by 2 observers and repeated 3 times by the primary observer. Results- Mean absolute differences between the single-plane fluoroscopic analysis and RSA measurements were 0.60, 1.28, and 0.64 mm for craniocaudal, proximodistal, and mediolateral translations, respectively, and 0.63°, 1.49°, and 1.58° for flexion-extension, abduction-adduction, and internal-external rotations, respectively. Intra- and interobserver repeatability was strong with maximum mean translational and rotational SDs of 0.52 mm and 1.36°, respectively. Conclusions and Clinical Relevance- Results suggested that single-plane fluoroscopic analysis performed by use of CT-derived bone models is a valid, noninvasive technique for accurately measuring 3-D femorotibial poses in dogs.

Objective- To assess kinetic 2-([(18)F]fluoro)-2-deoxy-d-glucose ((18)FDG) uptake in the brain of anesthetized healthy adult dogs by use of positron emission tomography (PET) and to determine whether (18)FDG uptake differs among anatomic regions of the brain. Animals- 5 healthy Beagles. Procedures- Each isoflurane-anesthetized dog was administered (18)FDG IV (dose range, 3.0 to 5.2 mCi), and PET data were acquired for 2 hours. A CT scan (without contrast agent administration) was performed to allow more precise neuroanatomic localization. Defined regions of interest within the brain were drawn on reconstructed image data. Standard uptake values (SUVs) for (18)FDG were calculated to generate time-activity curves and determine time to peak uptake. Results- Time-activity curve analysis identified 4 regional uptake patterns: olfactory, gray matter, white matter, and other (brainstem, cerebellum, and occipital and frontal regions). The highest maximum SUVs were identified in the olfactory bulbs and cerebral gray matter, and the lowest maximum SUV was identified in cerebral white matter. Mean time to peak uptake ranged from 37.8 minutes in white matter to 82.7 minutes in the olfactory bulbs. Conclusions and Clinical Relevance- Kinetic analysis of (18)FDG uptake revealed differences in uptake values among anatomic areas of the brain in dogs. These data provide a baseline for further investigation of (18)FDG uptake in dogs with immune-mediated inflammatory brain disease and suggest that (18)FDG-PET scanning has potential use for antemortem diagnosis without histologic analysis and for monitoring response to treatment. In clinical cases, a 1-hour period of PET scanning should provide sufficient pertinent data.

Objective-To determine associations of blood analysis variables and orbit and nasal planum surface temperatures with the onset and severity of Mycoplasma bovis pneumonia in calves. Animals-28 healthy calves. Procedures-Calves were challenged with M bovis (n = 24) on day 0 or not challenged (4). Blood samples were obtained for cardiac troponin I, CBC, and serum biochemical analyses on various days. Orbit and nasal planum surface temperatures were determined with infrared thermography on various days. Calves were euthanized, gross necropsies were performed, heart and lung samples were collected for histologic evaluation, and microbial cultures of lung samples were performed on day 14. Pneumonia severity was categorized as mild (< 10% lung consolidation) or moderate (≥ 10% lung consolidation). Associations between measured variables and severity of pneumonia or sample collection day were determined. Results-Plasma cardiac troponin I concentration for the 28 calves was significantly higher on day 14 than it was on day 0 or 7 (least squares mean, 0.02, 0, and 0 ng/mL, respectively). No other variables changed significantly during the study. No substantial gross or histologic abnormalities were identified in cardiac muscle samples. Day 14 plasma fibrinogen concentration was significantly different between calves with mild pneumonia and those with moderate pneumonia (mean, 0.44 and 0.74 g/dL, respectively). Calves with moderate pneumonia had significantly lower least squares mean surface temperature of the dorsal aspect of the nasal planum (18.7°C) versus calves with mild pneumonia (22.9°C). Conclusions and Clinical Relevance-Results indicated the evaluated variables had low value for assessment of bovine respiratory disease complex in calves.

Objective-To evaluate the cardiorespiratory effects of IV administration of propofol (4 mg/kg), ketamine hydrochloride and propofol (2 mg/kg each; K-P), or ketamine hydrochloride (5 mg/kg) and diazepam (0.2 mg/kg; K-D) before and after induction of anesthesia (IoA) in dogs sedated with acepromazine maleate and oxymorphone hydrochloride. Animals-10 healthy adult Beagles. Procedures-Each dog was randomly allocated to receive 2 of 3 treatments (1-week interval). For instrumentation prior to each treatment, each dog was anesthetized with isoflurane. After full recovery, acepromazine (0.02 mg/kg) and oxymorphone (0.05 mg/kg) were administered IV. Fifteen minutes later (before IoA), each dog received treatment IV with propofol, K-P, or K-D. Cardiorespiratory and arterial blood gas variables were assessed before, immediately after, and 5 minutes after IoA. Results-Compared with findings before IoA, dogs receiving the K-P or K-D treatment had increased cardiac output, oxygen delivery, and heart rate 5 minutes after IoA; K-P administration did not change mean arterial blood pressure or stroke volume and decreased systemic vascular resistance. Propofol decreased mean arterial blood pressure and systemic vascular resistance immediately after IoA but did not change heart rate, cardiac output, or oxygen delivery. All treatments caused some degree of apnea, hypoventilation, and hypoxemia (Pao2 < 80 mm Hg). Conclusions and Clinical Relevance-In dogs, K-P treatment maintained mean arterial blood pressure better than propofol alone and increased heart rate, cardiac output, or oxygen delivery, as did the K-D treatment. Supplemental 100% oxygen should be provided during IoA with all 3 treatments.

Objective-To ultrasonographically quantify experimentally induced effusion of the distal interphalangeal (DIP) joint of horses and compare results with those obtained with palpation. Sample-8 forelimbs from equine cadavers and forelimbs of 5 mares. Procedures-Preliminary ex vivo experiments were performed to validate the methods. Then, the DIP joints of the forelimbs of standing horses were serially distended with saline (0.9% NaCl) solution (1, 4, and 10 mL) by injection through an intra-articular catheter. Two ultrasonographers measured distension of the dorsal recess of the DIP joint, and 2 surgeons, who were not aware of the volume injected, graded joint effusion by palpation. Results-Intraobserver and interobserver repeatability was excellent for ultrasonographic measurements. Interobserver agreement for use of palpation to detect joint distension was moderate (κ = 0.45). There was an overall increase in the palpation distension grade with an increase in injected volume. Sensitivity for detection with palpation of larger volumes (4 and 10 mL) was high (92% and 100%, respectively). However, sensitivity was lower (57%) for detection with palpation of minimal distension (1 mL). Conclusions and Clinical Relevance-Although palpation provided a reliable clinical assessment of DIP joint effusion for volumes of 4 to 10 mL, ultrasonographic measurements were easy to obtain, more accurate, and able to detect smaller amounts of distension. This may be clinically relevant for the assessment of effusion of the DIP joint that can arise in horses with early osteoarthritis or infectious arthritis with concomitant soft tissue swelling that precludes accurate assessment with palpation.