Assessment of canine sensory function by using sine-wave electrical stimuli paradigm.

The paradigm of sine-wave electrical stimuli has been used for sensory neurological assessment in humans. In the present study, we applied the paradigm to the dog for the quantitative assessment of sensory function. Sine-wave electrical current stimuli at frequencies of 2000, 250, and 5Hz were delivered to bipolar electrodes attached to the skin surface of the hind paws. The stimulation intensity was gradually increased, and the minimum intensity required to elicit the lifting behavior in the stimulated paw was determined as current threshold (CT) for each of the three frequencies. Dogs consistently showed the lifting behavior at CTs without showing aversive behaviors such as vocalization and wriggling. The baseline CTs (mean+/-SEM, n=12) were 4430+/-110microA for CT2000, 2215+/-173microA for CT250, and 2305+/-152microA for CT5. The CTs immediately increased after bolus intravenous injection of fentanyl at 10microg/kg, although the significant increase disappeared within 1h. The time course for the CTs was parallel to that of plasma fentanyl concentration. In conclusion, the present study applied the paradigm of transcutaneous sine-wave electrical stimuli to the dog, and used the hind paw lifting as endpoint behavior. This paradigm is simple, non-invasive, useful in the assessment of sensory function, and can be adapted to investigate the pharmacokinetics/pharmacodynamics relation of drugs. Further studies are needed to give the conclusive interpretation of the endpoint behavior.

Antinociceptive and side-effects of hydromorphone after subcutaneous administration in cats.

The subcutaneous (SC) route is often chosen for drug administration in cats because it is easier to perform than intravenous (IV) injection and is perceived as less painful than intramuscular (IM) injection. However, little is known of how the route of administration influences the pharmacodynamics of drugs. This study measured the changes in skin temperature and thermal threshold (TT) and recorded the side-effects after SC injection of 0.1mg/kg of hydromorphone in six cats. Time to peak TT was 105min. Skin temperature was elevated at 15min and between 45 and 360min. Five cats vomited and two exhibited marked dysphoria. Compared to previously published studies of IV and IM administration of hydromorphone, the SC route results in a slower onset of peak effect, a shorter duration of antinociception and is associated with more undesirable side-effects. As with IV and IM injections, SC administration of hydromorphone at 0.1mg/kg is associated with a significant elevation in skin temperature. Overall, the SC route appears to have the least utility.

Development of a canine nociceptive thermal escape model.

Acute nociceptive models which have been validated for large animal species are limited, yet nociceptive assessment in non-rodent species is important in analgesic drug development where larger animals may be necessary because of the technical requirements of the study. Here we report development and validation of a canine hind paw thermal escape model and the effect of analgesics on withdrawal latencies. Individual focused projection bulbs were used as left and right voltage-adjusted thermal stimuli placed below a glass plate in a specifically designed canine holding apparatus. After acclimation, dogs were lightly restrained in a fabric sling while standing on the glass plate. The anterior center of the metatarsal pad of the left and right hind paw was positioned on the glass over each light, and duration of stimulation tolerance timed. For every trial, the escape latency from lamp actuation to paw withdrawal was recorded twice for each hind paw. The mean population baseline withdrawal latency of 9.3+/-1.7s (mean+/-S.D., n=12 dogs) was shown to be repeatable between paws, within and between individual animals, and between test days. This latency corresponded to a glass surface temperature of 49.5 degrees C. A cut-off time of 20s (corresponding to a glass surface temperature of 56.5 degrees C) was set to prevent tissue damage. Intravenous administration (mg/kg) of morphine (1.0), hydromorphone (0.2), butorphanol (0.4), fentanyl (0.01), and dexmedetomidine (0.01) significantly (p<0.05) increased withdrawal latency from baseline within 15-30 min of administration while buprenorphine (0.03) produced a delayed, modest but significant latency increase. Rank order of opioid analgesic duration was morphine=hydromorphone>butorphanol>bupenorphine>fentanyl=saline. A dose-effect curve for hydromorphone was generated and corresponded to previously described dose-effect relationships in other species. The non-analgesic tranquilizer acepromazine (0.1mg/kg) produced mild sedation, but no significant increase in latency from that of saline. The model yielded a clear distinction between analgesia and sedation for all agents tested. These studies provide validation of a canine thermal escape model and have demonstrated the efficacy of clinically relevant doses of analgesics in elevating escape latencies. This model will facilitate quantification of the effects of parenterally and neuraxially administered analgesics in dogs.

The quantitation of procaine in equine plasma by liquid chromatography-linear ion trap mass spectrometry.

A method for the extraction and quantitation of procaine in equine plasma was developed for use with liquid chromatography-mass spectrometry (LC-MS). Procaine was isolated from equine plasma by liquid-liquid extraction at pH 11 with dichloromethane using procaine-d10 as an internal standard. Quantitation was achieved by LC-MS using a 3-microm C-18 column coupled to an electrospray ionization source on a linear ion-trap mass spectrometer. The limit of detection and limit of quantitation was determined to be 50 and 200 pg/mL, respectively. The lowest limit of detection determined by previous methods was 1 ng/mL. Administration samples were obtained as part of a larger study to determine a regulatory limit for procaine in racehorses and procaine concentrations were determined using this method.

Plasma concentration and local anesthetic activity of procaine hydrochloride following subcutaneous administration to horses.

OBJECTIVE: To determine the durations of the local anesthetic effect and plasma procaine concentrations associated with 5- and 10-mg doses of procaine hydrochloride (with or without 100 microg of epinephrine) administered SC over the lateral palmar digital nerves of horses. ANIMALS: 6 healthy adult horses. PROCEDURES: The hoof withdrawal reflex latency (HWRL) period was determined by use of a focused heat lamp before and after administration of procaine with and without epinephrine. Blood samples were collected immediately before determination of each HWRL period to assess plasma concentrations of procaine via liquid chromatography-mass spectrometry-mass spectrometry (LC-MS-MS). RESULTS: 10 but not 5 mg of procaine alone and 5 and 10 mg of procaine administered with epinephrine significantly prolonged the HWRL period (mean durations of effect, 5, 120 and 180 minutes, respectively), compared with baseline values. Plasma procaine concentrations did not correlate well with local anesthetic activity; for example, although the HWRL was prolonged to the maximum permitted duration of 20 seconds at 60 to 180 minutes following administration of the 5-mg dose of procaine with epinephrine in certain horses, plasma procaine concentrations were less than the limit of quantitation of the LC-MS-MS assay. CONCLUSIONS AND CLINICAL RELEVANCE: Small doses of procaine coadministered with epinephrine provided long-lasting local analgesia and resulted in plasma procaine concentrations that were not always detectable via LC-MS-MS. On the basis of these results, the use of regulatory limits or thresholds for procaine concentration in equine plasma samples obtained after racing should be seriously reconsidered.

Dose-related thermal antinociceptive effects of intravenous hydromorphone in cats.

OBJECTIVE: To describe the dose-related thermal antinociceptive effects of intravenous (i.v.) hydromorphone in cats. STUDY DESIGN: Randomized, blinded, crossover design. ANIMALS: Seven adult cats (3.5-7.4 kg), two spayed females, and five neutered males. METHODS: Hydromorphone (0.025, 0.05, or 0.1 mg kg(-1)) was administered i.v.. Skin temperature and thermal threshold were measured before and at selected time points to 720 minutes post-administration. Statistical analysis of mean thermal threshold and skin temperatures over time for each dose and between doses was by way of a split-plot model and post hoc Bonferroni t-tests. p < 0.05 was considered significant. RESULTS: A significant difference from baseline for mean thermal threshold was identified for the 0.05 mg kg(-1) dose (5-80 minutes, peak thermal threshold 46.9 +/- 6.2 degrees C) and 0.1 mg kg(-1) dose (5-200 minutes, peak thermal threshold 54.9 +/-0.2 degrees C). The thermal threshold was significantly greater after the 0.1 mg kg(-1) dose from 5 to 200 minutes compared to the 0.025 mg kg(-1) and 0.5 mg kg(-1) doses. The thermal threshold was significantly greater from 35 to 80 minutes for the 0.05 mg kg(-1) dose when compared with the 0.025 mg kg(-1) dose. Skin temperature was significantly increased from 35 to 140 minutes following the 0.1 mg kg(-1) dose. CONCLUSIONS: A dose-related antinociceptive effect was demonstrated for i.v. hydromorphone in cats. CLINICAL RELEVANCE: Hydromorphone at doses less than 0.1 mg kg(-1) has a modest antinociceptive effect and a short duration of action. At a dose of 0.1 mg kg(-1) i.v., onset of analgesia is rapid with a clinically useful duration of effect, but is associated with a rise in skin temperature.

Time course and role of morphine dose and concentration in intrathecal granuloma formation in dogs: a combined magnetic resonance imaging and histopathology investigation.

BACKGROUND: Intrathecal morphine infusion leads to intrathecal granulomas. In dogs, the authors examined time course of granuloma formation and the role of concentration in granuloma development. METHODS: Dogs were prepared with lumbar intrathecal catheters and vest-mounted pumps. To define the time course of granuloma formation, serial magnetic resonance imaging was performed in animals receiving 10 or 31 days of morphine infusion (12.5 mg/ml at 40 microl/h). At these times, morphine was removed from the infusate, and further magnetic resonance images were acquired over 14-35 additional days. To assess dose versus concentration, dogs received 28-day infusions of vehicle, 12 mg morphine/day as 12.5 mg/ml at 40 microl/h, or 1.5 mg/ml at 334 microl/h (12 mg/day) for 28 days. Additional dogs received 3 mg/day as 12.5 mg/ml at 10 mul/h. RESULTS: Serial magnetic resonance images in dogs receiving morphine (12.5 mg/ml at 40 microl/h) revealed pericatheter-enhancing tissues as early as 3 days with a prominent signal by 10 days. Removal of morphine reduced the mass volume within 7 days. At a fixed infusion rate, the incidence of granuloma formation with the continuous intrathecal infusion of morphine ranged from 0 in vehicle-treated dogs to 100% in dogs treated with 12.5 mg/ml at 40 microl/h (12 mg/day). Infusion of 12 mg/day at 1.5 mg/ml (334 microl/h) resulted in granuloma in one of four animals. The authors found that infusion of morphine in different concentrations at a fixed rate resulted in a dose-dependent increase in concentration, with the granuloma-producing, dose-yielding lumbar cerebrospinal fluid morphine concentrations around 40 microg/ml. CONCLUSIONS: Serial magnetic resonance imaging showed a rapid formation and regression of the masses initiated by intrathecal morphine infusion. These masses are dependent on local concentration.