Long-acting injectable methadone (methadone-fluconazole) provides safe and effective postoperative analgesia in a randomized clinical trial for dogs undergoing soft tissue surgery.

OBJECTIVE: To assess the pharmacokinetics, clinical efficacy, and adverse effects of injectable methadone with the pharmacokinetic enhancer fluconazole (methadone-fluconazole), compared with the standard formulation of injectable methadone, in dogs after ovariohysterectomy. We hypothesized that 2 doses of methadone-fluconazole would provide 24 hours of postoperative analgesia. ANIMALS: 3 purpose-bred dogs (pharmacokinetic preliminary study) and 42 female dogs from local shelters (clinical trial) were included. PROCEDURES: Pharmacokinetics were preliminarily determined. Clinical trial client-owned dogs were blocked by body weight into treatment groups: standard methadone group (methadone standard formulation, 0.5 mg/kg, SC, q 4 h; n = 20) or methadone-fluconazole group (0.5 mg/kg methadone with 2.5 mg/kg fluconazole, SC, repeated once at 6 h; n = 22). All dogs also received acepromazine, propofol, and isoflurane. Surgeries were performed by experienced surgeons, and dogs were monitored perioperatively using the Glasgow Composite Measure Pain Scale-Short Form (CMPS-SF) and sedation scales. Evaluators were masked to treatment. RESULTS: Findings from pharmacokinetic preliminary studies supported that 2 doses of methadone-fluconazole provide 24 hours of drug exposure. The clinical trial had no significant differences in treatment failures or postoperative CMPS-SF scores between treatments. One dog (methadone-fluconazole group) had CMPS-SF > 6 and received rescue analgesia. All dogs had moderate sedation or less by 1 hour (methadone-fluconazole group) or 4 hours (standard methadone group) postoperatively. Sedation was completely resolved in all dogs the day after surgery. CLINICAL RELEVANCE: Methadone-fluconazole with twice-daily administration was well tolerated and provided effective postoperative analgesia for dogs undergoing ovariohysterectomy. Clinical compliance and postoperative pain control may improve with an effective twice-daily formulation.

Assessment of agreement among diplomates of the American College of Veterinary Anesthesia and Analgesia for scoring the recovery of horses from anesthesia by use of subjective grading scales and development of a system for evaluation of the recovery of horses from anesthesia by use of accelerometry.

OBJECTIVE To evaluate agreement among diplomates of the American College of Veterinary Anesthesia and Analgesia for scores determined by use of a simple descriptive scale (SDS) or a composite grading scale (CGS) for quality of recovery of horses from anesthesia and to investigate use of 3-axis accelerometry (3AA) for objective evaluation of recovery. ANIMALS 12 healthy adult horses. PROCEDURES Horses were fitted with a 3AA device and then were anesthetized. Eight diplomates evaluated recovery by use of an SDS, and 7 other diplomates evaluated recovery by use of a CGS. Agreement was tested with κ and AC1 statistics for the SDS and an ANOVA for the CGS. A library of mathematical models was used to map 3AA data against CGS scores. RESULTS Agreement among diplomates using the SDS was slight (κ = 0.19; AC1 = 0.22). The CGS scores differed significantly among diplomates. Best fit of 3AA data against CGS scores yielded the following equation: RS = 9.998 × SG0.633 × ∑UG0.174, where RS is a horse's recovery score determined with 3AA, SG is acceleration of the successful attempt to stand, and ∑UG is the sum of accelerations of unsuccessful attempts to stand. CONCLUSIONS AND CLINICAL RELEVANCE Subjective scoring of recovery of horses from anesthesia resulted in poor agreement among diplomates. Subjective scoring may lead to differences in conclusions about recovery quality; thus, there is a need for an objective scoring method. The 3AA system removed subjective bias in evaluations of recovery of horses and warrants further study.

Effects of lidocaine administration via continuous rate infusion on the minimum alveolar concentration of isoflurane in New Zealand White rabbits (Oryctolagus cuniculus).

OBJECTIVE: To evaluate the effect of a continuous rate infusion (CRI) of lidocaine on the minimum alveolar concentration (MAC) of isoflurane in rabbits. ANIMALS: Five 12-month-old female New Zealand White rabbits (Oryctolagus cuniculus). PROCEDURES: Rabbits were anesthetized with isoflurane. Baseline isoflurane MAC was determined by use of the tail clamp technique. A loading dose of lidocaine (2.0 mg/kg, IV) was administered followed by a CRI of lidocaine at 50 µg/kg/min. After 30 minutes, isoflurane MAC was determined. Another loading dose was administered, and the lidocaine CRI then was increased to 100 µg/kg/min. After 30 minutes, isoflurane MAC was determined again. Plasma samples were obtained for lidocaine analysis after each MAC determination. RESULTS: Baseline isoflurane MAC was 2.09%, which was similar to previously reported values in this species. Lidocaine CRI at 50 and 100 µg/kg/min induced significant reductions in MAC. The 50 µg/kg/min CRI resulted in a mean plasma lidocaine concentration of 0.654 µg/mL and reduction of MAC by 10.5%. The 100 µg/kg/min CRI of lidocaine resulted in a mean plasma concentration of 1.578 µg/mL and reduction of MAC by 21.7%. Lidocaine also induced significant decreases in arterial blood pressure and heart rate. All cardiopulmonary variables were within reference ranges for rabbits anesthetized with inhalation anesthetics. No adverse effects were detected; all rabbits had an uncomplicated recovery from anesthesia. CONCLUSIONS AND CLINICAL RELEVANCE: Lidocaine administered as a CRI at 50 and 100 µg/kg/min decreased isoflurane MAC in rabbits. The IV administration of lidocaine may be a useful adjunct in anesthesia of rabbits.

Use of computed tomography to evaluate pathologic changes in the lungs of calves with experimentally induced respiratory tract disease.

OBJECTIVE: To optimize methods for the use of computed tomography (CT) to assess pathologic changes in the lungs of calves and to determine the effect of treatment on lung consolidation. ANIMALS: 10 male Holstein calves. PROCEDURES: Calves were anesthetized to facilitate CT imaging of the thorax. After initial images were obtained, pneumonia was induced in the calves by inoculation through a bronchoscope. Two calves were used in a preliminary study to refine the inoculation dose and optimize CT images. Four calves were administered florfenicol and 4 calves were untreated control animals. Serial images were obtained 24, 48, and 72 hours after inoculation. After final images were obtained, calves were euthanized, and lung consolidation was estimated by use of lung surface area scoring and water displacement. These estimates were compared with estimated lung consolidation obtained by use of CT. RESULTS: Calves had rapid disease progression. Percentage of lung consolidation was not significantly different between treatment groups for any of the estimation methods. Results of an ANOVA of the 3 assessment methods indicated significant differences among methods. Estimates of the percentage of lung consolidation obtained by use of surface area scoring and CT correlated well, whereas water displacement estimates correlated poorly with other methods of consolidation estimation. CONCLUSIONS AND CLINICAL RELEVANCE: Because of the correlation with other methods for estimation of lung consolidation, CT has the potential to be used to monitor disease progression in calves with experimentally induced respiratory tract disease.

Comparison of radiographic and computed tomography lymphangiography for identification of the canine thoracic duct.

Standard radiographic lymphangiograms and computed tomography (CT) lymphangiograms were performed on 10 female dogs without intrathoracic disease. Positive contrast lymphagiography was performed by injection into a catheterized mesenteric lymphatic vessel, and lateral thoracic radiographs, ventrodorsal thoracic radiographs, and thoracic CTs were obtained. The number of visible ducts was recorded for each image at the midbody of the ninth thoracic vertebra (T9) through the first lumbar vertebra (L1). Data were combined for all dogs at each data acquisition point. Data were analyzed by comparing data from all three images independently, and then by combining data for the radiographs and comparing the study with the highest number of visible duct branches to the CT. Significant differences in numbers of branches were found at T11 and L1. This study suggests that CT may be able to quantify branches of the thoracic duct more accurately than standard radiographic lymphangiography.

Use of a continuous, local infusion of bupivacaine for postoperative analgesia in dogs undergoing total ear canal ablation.

OBJECTIVE: To determine whether addition of a continuous, local infusion of bupivacaine would improve postoperative analgesia in dogs undergoing total ear canal ablation. DESIGN: Randomized controlled trial. ANIMALS: 16 dogs undergoing total ear canal ablation (12 unilaterally and 4 bilaterally with > 1 month between procedures). PROCEDURE: Dogs were randomly allocated to receive morphine (0.25 mg/kg [0.11 mg/lb]) at the end of the procedure (10 procedures) or morphine and a continuous, local infusion of bupivacaine (0.13 to 0.21 mg/kg/h [0.06 to 0.1 mg/lb/h]; 10 procedures). Dogs were observed for 48 hours after surgery. Additional doses of morphine were administered up to every 4 hours in dogs with signs of severe pain. RESULTS: Temperament, sedation, analgesia, and cumulative pain scores were not significantly different between groups any time after surgery. Recovery score was significantly higher for dogs that received bupivacaine than for control dogs 2 hours after extubation but not at any other time. Serum cortisol concentration was not significantly different between groups at any time but, in both groups, was significantly increased at the time of extubation, compared with all other observation times. Total number of additional doses of morphine administered was not significantly different between groups. Bupivacaine was not detected in the plasma of any of the dogs that received the local bupivacaine infusion. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that addition of a continuous, local infusion of bupivacaine did not significantly increase the degree of postoperative analgesia in dogs that underwent total ear canal ablation and were given morphine at the end of surgery.

Transnasal laryngoscopy for the diagnosis of laryngeal paralysis in dogs.

Four dogs with clinical signs of laryngeal paralysis and three normal dogs were evaluated with transnasal laryngoscopy. Six of these dogs subsequently underwent standard laryngoscopy. For transnasal laryngoscopy, a video endoscope was passed through the left nasal passage after intramuscular sedation and topical anesthesia. The laryngeal opening was observed during spontaneous ventilation. Laryngeal paralysis was diagnosed in four dogs and was confirmed with traditional laryngoscopy in three dogs. Normal motion of the arytenoid cartilages was present in the other three dogs; however, two required mechanical stimulation of the laryngeal mucosa for full evaluation. Transnasal laryngoscopy provided a means for diagnosing laryngeal paralysis in dogs without general anesthesia.

Popliteal and mesenteric lymph node injection with methylene blue for coloration of the thoracic duct in dogs.

OBJECTIVE: To describe and compare the time of onset and intensity of thoracic duct coloration after injection of methylene blue into a mesenteric or popliteal lymph node. STUDY DESIGN: Experimental study. ANIMALS: Twenty adult dogs. METHODS: A right tenth intercostal thoracotomy, a right paracostal laparotomy, and an approach to the right popliteal lymph node were performed on each dog. Methylene blue (0.5 mg/kg of a 1% solution, maximum 10 mg) was injected into either a mesenteric (group M, 10 dogs) or popliteal (group P, 10 dogs) lymph node. Thoracic duct color was graded (0 to 3) every 5 minutes for 60 minutes. Statistical analysis was performed on mean thoracic duct color grade data, on number of successful outcomes between groups M and P, and between weight groups. RESULTS: Coloration of the thoracic duct occurred in all group M dogs and 6 group P dogs. Coloration was first recorded 0 to 10 minutes after injection in all dogs and persisted for 60 minutes in 15 dogs. Mean thoracic duct color grade was significantly increased postinjection compared with preinjection at all times in group M. More successful outcomes occurred in group M (P =.03). CONCLUSIONS: Methylene blue injected into mesenteric or popliteal lymph nodes was successful in coloring the thoracic duct, but both mean grade and number of successful outcomes were significantly higher after mesenteric injection. CLINICAL RELEVANCE: Thoracic duct coloration after lymph node injection occurred within 10 minutes and persisted for 60 minutes. This information is useful in planning thoracic duct ligation in cases of chylothorax when observation of the duct is desired. Injection of both lymph node sites was successful, but mesenteric node injection was a more reliable technique.

4-aminopyridine decreases progesterone production by porcine granulosa cells.

BACKGROUND: Ion channels occur as large families of related genes with cell-specific expression patterns. Granulosa cells have been shown to express voltage-gated potassium channels from more than one family. The purpose of this study was to determine the effects of 4-aminopyridine (4-AP), an antagonist of KCNA but not KCNQ channels. METHODS: Granulosa cells were isolated from pig follicles and cultured with 4-AP, alone or in combination with FSH, 8-CPT-cAMP, estradiol 17beta, and DIDS. Complimentary experiments determined the effects of 4-AP on the spontaneously established pig granulosa cell line PGC-2. Granulosa cell or PGC-2 function was assessed by radio-immunoassay of media progesterone accumulation. Cell viability was assessed by trypan blue exclusion. Drug-induced changes in cell membrane potential and intracellular potassium concentration were documented by spectrophotometric determination of DiBAC4(3) and PBFI fluorescence, respectively. Expression of proliferating cell nuclear antigen (PCNA) and steroidogenic acute regulatory protein (StAR) was assessed by immunoblotting. Flow cytometry was also used to examine granulosa cell viability and size. RESULTS: 4-AP (2 mM) decreased progesterone accumulation in the media of serum-supplemented and serum-free granulosa cultures, but inhibited cell proliferation only under serum-free conditions. 4-AP decreased the expression of StAR, the production of cAMP and the synthesis of estradiol by PGC-2. Addition of either 8-CPT-cAMP or estradiol 17beta to serum-supplemented primary cultures reduced the inhibitory effects of 4-AP. 4-AP treatment was also associated with increased cell size, increased intracellular potassium concentration, and hyperpolarization of resting membrane potential. The drug-induced hyperpolarization of resting membrane potential was prevented either by decreasing extracellular chloride or by adding DIDS to the media. DIDS also prevented 4-AP inhibition of progesterone production. CONCLUSION: 4-AP inhibits basal and FSH-stimulated progesterone production by pig granulosa cells via drug action at multiple interacting steps in the steroidogenic pathway. These inhibitory effects of 4-AP on steroidogenesis may reflect drug-induced changes in intracellular concentrations of K+and Cl- as well as granulosa cell resting membrane potential.

Potassium channel antagonists influence porcine granulosa cell proliferation, differentiation, and apoptosis.

This investigation determined the effects of K(+) channel antagonists on proliferation, differentiation, and apoptosis of porcine granulosa cells. The drugs screened for functional effects included the class III antiarrhythmic agents MK-499 and clofilium, the chromanol I(Ks) antagonist 293B, the benzodiazepine I(Ks) antagonists L-735,821 and L-768,673, and the peptidyl toxins charybdotoxin (CTX) and margatoxin (MTX). Granulosa cell proliferation and differentiation were assessed by serial measurements of cell number and progesterone accumulation in the culture media, respectively. Granulosa cell apoptosis was evaluated using flow cytometry. Additional information about drug effects was obtained by immunoblotting to detect expression of proliferating cell nuclear antigen, p27(kip1) and the caspase-3 substrate poly(ADP-ribose) polymerase. The ERG channel antagonist MK-499 had no functional effects on cultured granulosa cells. However, the broad spectrum K(+) channel antagonist clofilium decreased, in a concentration-dependent fashion, the number of viable granulosa cells cultured, and these effects were associated with induction of apoptosis. All three I(Ks) antagonists (293B, L-735,821, and L-768,673) increased basal, but not FSH-enhanced progesterone accumulation on Day 1 after treatment without affecting the number of viable cells in culture, an effect that was blocked by pimozide. In contrast, CTX and MTX increased the number of viable cells in FSH-stimulated cultures on Day 3 after treatment without affecting progesterone output per cell. These data demonstrate that selective antagonism of granulosa cell K(+) channels with distinct molecular correlates, electrophysiological properties, and expression patterns can influence differential granulosa cell proliferation, steroidogenic capability, and apoptosis. Thus, K(+) channels may represent pharmacological targets for affecting Granulosa cell function and oocyte maturation, in vivo or in vitro.

Thoracoscopic visualization and ligation of the thoracic duct in dogs.

OBJECTIVE: To develop a technique for thoracoscopic visualization and ligation of the thoracic duct in dogs. STUDY DESIGN: In vivo experimental study. ANIMALS: Five mature, healthy dogs. METHODS: Dogs were normal based on physical examination, negative occult heartworm test, normal complete blood count and biochemical profile, and normal thoracic radiographs. The dogs were anesthetized, and a ventral midline laparotomy was performed for catheterization of a mesenteric lymphatic. Lymphangiography was performed to determine thoracic duct anatomy. Thoracoscopy was performed in the caudal, right hemithorax after single lung intubation or bronchial blockade. At least two 10-mm clips were placed across the thoracic duct in each dog. Lymphangiography was repeated to assess duct ligation. If complete duct occlusion was not achieved, thoracoscopy was repeated for additional clip placement. After surgery the dogs were euthanatized, and necropsies were performed. RESULTS: Lymphangiography showed that multiple branches of the thoracic duct were present in every dog; bilateral thoracic duct branches were most common. Thoracoscopic identification and ligation of the thoracic duct was successful in all five dogs. Two dogs required a second thoracoscopic procedure to completely occlude flow of contrast through the thoracic duct. Surgery time for thoracoscopy averaged 59 plus minus 9.6 minutes. Retroperitoneal contrast accumulation after thoracic duct ligation occurred in two dogs. One dog required bilateral pulmonary ventilation. CONCLUSION: Thoracoscopy can be used to visualize the thoracic duct for ligation in normal dogs. CLINICAL RELEVANCE: Thoracoscopic ligation of the thoracic duct may be a therapeutic option for management of chylothorax in dogs.

Molecular basis of voltage-dependent potassium currents in porcine granulosa cells.

The major objective of this study was to elucidate the molecular bases for K(+) current diversity in porcine granulosa cells (GC). Two delayed rectifier K(+) currents with distinct electrophysiological and pharmacological properties were recorded from porcine GC by using whole-cell patch clamp: 1) a slowly activating, noninactivating current (I(Ks)) antagonized by clofilium, 293B, L-735,821, and L-768,673; and 2) an ultrarapidly activating, slowly inactivating current (I(Kur)) antagonized completely by clofilium and 4-aminopyridine and partially by tetraethylammonium, charybdotoxin, dendrotoxin, and kaliotoxin. The molecular identity of the K(+) channel genes underlying I(Ks) and I(Kur) was examined using reverse transcription-polymerase chain reaction and immunoblotting to detect K(+) channel transcripts and proteins. We found that GC could express multiple voltage-dependent K(+) (Kv) channel subunits, including KCNQ1, KCNE1, Kv1.1, Kv1.2, Kv1.3, Kv1.4, Kv1.5, Kv1.6, Kvbeta1.3, and Kvbeta2. Coimmunoprecipitation was used to establish the hetero-oligomeric nature of granulosa cell Kv channels. KCNE1 and KCNQ1 were coassociated in GC, and their expression coincided with the expression of I(Ks). Extensive coassociation of the various Kv alpha- and beta-subunits was also documented, suggesting that the diverse electrophysiological and pharmacological properties of I(Kur) currents may reflect variation in the composition and stoichiometry of the channel assemblies, as well as differences in post-translational modification of contributing Kv channel subunits. Our findings provide an essential background for experimental definition of granulosa K(+) channel function(s). It will be critical to define the functional roles of specific GC K(+) channels, because these proteins may represent either novel targets for assisted reproduction or potential sites of drug toxicity.