Some azo dyes, including Sudans I–IV and Para Red, are genotoxic and may be biotransformed to cancerogenic aromatic amines. They are banned as food and feed additives, but their presence has been detected in food. Aromatic amines are also considered potentially toxic. Online EC–MS is a promising tool to study the transformation mechanisms of xenobiotics such as azo dyes. The aim of the study was to investigate emulation of how azo dyes are enzymatically transformed to amines with EC–MS. The reduction reactions of five azo dyes (Sudans I–IV and Para Red) were conducted using a glassy carbon working electrode and 0.1% formic acid in acetonitrile. Reduction results were compared with the literature and in silico to select preliminary candidates for metabolites. The LC-MS/MS method was used to confirm results obtained by electrochemical reactor. A limited number of pre-selected compounds were confirmed as azo dyes metabolites – aniline for Sudan I, aniline and 4-aminoazobenzene for Sudan III, o-toluidine for Sudan IV, and 4-nitroaniline for Para Red. No metabolites were found for Sudan II. Electrochemistry–mass spectrometry was successfully applied to azo dyes. This approach may be used to mimic the metabolism of azo dyes, and therefore predict products of biotransformation.

OBJECTIVE To determine effects of catheter shape, interelectrode spacing (IS), and electrode size (ES) on pacing threshold (PT), extraneous muscular stimulation (EMS), and zone of capture (ZOC) for dogs undergoing transesophageal atrial pacing (TAP). ANIMALS 10 purpose-bred dogs without cardiac conduction disturbances. PROCEDURES 7 configurations for TAP catheters were tested in each dog to evaluate effects of catheter shape (curved or straight), IS (5, 15, and 25 mm), and ES (2, 4, and 6 mm). Each catheter was passed into the esophagus to a location aboral to the heart and slowly withdrawn until atrial pacing was achieved. Then, catheters were withdrawn in 5-mm increments until pacing could not be achieved. Outcomes measured at each pacing site included PT, degree of EMS, and ZOC. RESULTS There was a significantly lower PT, wider ZOC, and less EMS for the curved catheter than for the straight catheter. An ES of 6 mm induced significantly more EMS than was induced by an ES of 2 or 4 mm. An IS of 5 mm induced significantly less EMS and a significantly narrower ZOC but required a significantly higher PT, compared with results for an ES of 15 or 25 mm. Additionally, there was a significant direct correlation between IS and ZOC. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that a curved catheter with multiple 4-mm electrodes that provides for variable IS would be ideal for TAP in dogs. IMPACT FOR HUMAN MEDICINE TAP catheters currently used in human medicine are straight. The PT in humans may potentially be reduced with curved catheters.

Owing to technical and ethical limitations, a substantial part of the knowledge about the pathophysiologic mechanism of the human diaphragm has been obtained from studies in which phrenic nerve activation was usually carried out by direct surgical exposure of the nerves in the neck of deeply anesthetized, mechanically ventilated animals. Novel information has been gleaned from such studies, but the restrictive conditions under which it was collected preclude reliable extrapolation. We, therefore, addressed the question of whether accurate electrophysiologic evaluation of the phrenic nerve-diaphragm pathway can be performed in intact, nonanesthetized calves. Transjugular phrenic activation was well tolerated, safe, specific, and able to achieve constant symmetric and supramaximal phrenic stimulations during prolonged periods. Eighteen noninvasive cutaneous and esophageal reception circuits were tested for their ability to record the diaphragmatic evoked potential. In addition, they were compared for specificity and reproducibility of the recorded potentials during prolonged periods of tidal or stimulated respiration. The best diaphragmatic potential was recorded from surface electrodes attached to the skin of the ninth and tenth intercostal spaces, using a xyphoidian reference. We describe a method that allows easy, long-term, and reliable electrophysiologic evaluation of the phrenic nerve-diaphragm pathway in intact, conscious calves. It is hoped that such a model will produce relevant novel information regarding pathophysiology of the diaphragm.

Zoometric measurements and bioelectrical impedance analysis were evaluated as methods of body composition determination in healthy cats. Zoometric and impedance measurements were taken on 22 anesthetized adult cats of various ages, genders, breeds, and body weights. The cats were then euthanatized. The bodies were processed through a tissue homogenizer and free-catch specimens were taken, freeze-dried, and analyzed for total body water, protein, fat, potassium, and ash content. Stepwise regression analysis was implemented to identify statistically significant relationships between the chemically determined dependent variables (total body water, protein, potassium, fat-free mass, fat mass, and percent body fat) and the zoometric measurements, with or without bioelectrical impedance analysis. Statistical analysis revealed high correlations between the dependent variables and the corresponding predicted values of those variables. Body weight alone was a poor predictor of body composition in these cats. On the basis of these findings, we suggest that zoometric and bioelectrical impedance measurements may serve as practical, noninvasive, simple, and accurate methods for estimating body composition in domestic cats.

Evaluation of urethral pressure profilometry (UPP) with simultaneous fine-wire electromyography of the external urethral sphincter (EUS) was conducted in 11 healthy adult male cats sedated with xylazine and ketamine. A 3.5-F urethral catheter with a closed end and two 1-mm sideports was infused with sterile 0.9% NaCl solution at a rate of 2 to 3 ml/min. A fine-wire electromyographic (EMG) electrode was placed percutaneously into or near the external urethral sphincter prior to the onset of the UPP. The maximal urethral pressure achieved and functional profile length were recorded from UPP. Setting both catheter withdrawal rate and paper speed at 5 mm/s enabled the measurement of actual urethral length directly from UPP. Sphincter EMG activity was rated as slight (+), moderate (+ +), or intense (+ + +). All recordings were replicated once during each trial for 8 cats and trials were replicated 5 to 7 days later in 4 cats. Before catheterization, EMG activity of the external urethral sphincter was rated slight (+), whereas intense (+ + +) activity accompanied insertion. The activity evoked by movement of the catheter subsided, but intense EMG activity of the external urethral sphincter was recorded from onset to completion of catheter withdrawal in all cats in both trials. The mean maximal urethral pressure was 93.1 +/- 13.29 cm H2O. The mean function urethral length was 8.1 +/- 0.93 cm. Maximal urethral pressure or function profile length did not differ significantly between recordings within trials or between trials. Simultaneous recording of EMG activity and UPP of the external urethral sphincter was shown to be a simple, noninvasive technique for assessing neuromuscular and anatomic urethral function.

Corynebacterium pseudotuberculosis was transformed by electroporation, using pNG2, an erythromycin-resistance plasmid from C diphtheriae. Corynebacterium pseudotuberculosis cultivated in brain-heart infusion broth was washed 3 times with water, and resuspended to a final concentration of about 5 X 10(13) colony-forming units/ml. An electroporator constructed in our laboratory incorporated an electrode with 0.8-mm interelectrode gap, using disposable spectrophotometer cuvettes as containers for electroporation. The pNG2 was prepared in Escherichia coli and 4 to 16 microgram of pNG2 DNA was mixed with 400-microliter amounts of cell suspension in prechilled cuvettes. After incubation on ice for 5 to 10 minutes, the mixture was electroporated at field strengths of up to 18 kV/cm, mixed with 1.5 ml of brain-heart infusion broth, and incubated at 37 C for 2 hours with agitation. Aliquots were then plated on brain-heart infusion blood agar with 15 microgram of erythromycin/ml. Corynebacterium pseudotuberculosis was transformed at a maximal efficiency of approximately 4 X 10(4) transformants/microgram of pNG2 DNA. Most total transformants and most transformants per microgram of pNG2 were generated at a field strength of 18 kV/cm. When the concentration of pNG2 DNA was varied, the average total number of transformants increased through a concentration of 30 microgram/ml, but the efficiency of transformation was highest at the lowest DNA concentration. Transformants contained unmodified pNG2.

Evoked potentials were induced by transcranial stimulation and recovered from the spinal cord, and the radial and sciatic nerves in six dogs. Stimulation was accomplished with an anode placed on the skin over the area of the motor cortex. Evoked potentials were recovered from the thoracic and lumbar spinal cord by electrodes placed transcutaneously in the ligamentum flavum. Evoked potentials were recovered from the radial and sciatic nerves by surgical exposure and electrodes placed in the perineurium. Signals from 100 repetitive stimuli were averaged and analyzed. Waveforms were analyzed for amplitude and latency. Conduction velocities were estimated from wave latencies and distance traveled. The technique allowed recovery of evoked potentials that had similar characteristics among all dogs. Conduction velocities of potentials recovered from the radial and sciatic nerves suggested stimulation of motor pathways; however, the exact origin and pathway of these waves is unknown.

Scalp electrode impedance measurements recorded by wired and wireless electroencephalography (EEG) machines in 7 healthy dogs were compared. Eight recordings resulted in 80 impedance readings from subdermal wire electrodes (locations F7/F8, F3/F4, T3/T4, C3/C4, Fz, and Cz). Impedance values were measured first from the wired and then the wireless EEG machine. Wireless impedance measurements were higher than the wired EEG machine in 79/80 readings (P ≤ 0.05), being on average 2.83 kΩ [P ≤ 0.05, 95% confidence interval (CI): 2.51 to 3.14, SD = 1.42] higher. Impedances from the wired machine ranged between < 0.5 and 9 kΩ (mean = 3.09, median = 2.00, SD = 2.15), whereas impedances from the wireless machine ranged between 2.69 and 6.07 kΩ (mean = 5.92, median = 5.05, SD = 2.59). Despite these differences in impedance measurements, both machines measured similar impedance patterns. The wireless EEG machine's impedance measurements, therefore, should be acceptable for veterinary clinical settings.

OBJECTIVE To determine the effect of oral administration of gabapentin (20 mg/kg) on the minimum alveolar concentration (MAC) of isoflurane in dogs. ANIMALS 6 healthy adult dogs (3 males and 3 females with a mean ± SD body weight of 24.8 ± 1.3 kg). PROCEDURES Each dog was anesthetized twice. Dogs were initially assigned to 1 of 2 treatments (gabapentin [20 mg/kg, PO] followed 2 hours later by anesthesia maintained with isoflurane or anesthesia maintained with isoflurane alone). A minimum of 7 days later, dogs received the other treatment. The MAC of isoflurane was determined by use of an iterative bracketing technique with stimulating electrodes placed in the maxillary buccal mucosa. Hemodynamic variables and vital parameters were recorded at the lowest end-tidal isoflurane concentration at which dogs did not respond to the stimulus. Effect of treatment on outcome variables was analyzed by use of a paired t test. RESULTS Mean ± SD MAC of isoflurane was significantly lower when dogs received gabapentin and isoflurane (0.71 ± 0.12%) than when dogs received isoflurane alone (0.91 ± 0.26%). Mean reduction in MAC of isoflurane was 20 ± 14%. Hemodynamic variables did not differ significantly between treatments. Mean time to extubation was significantly less when dogs received gabapentin and isoflurane (6 ± 4 minutes) than when dogs received isoflurane alone (23 ± 15 minutes). CONCLUSIONS AND CLINICAL RELEVANCE Oral administration of gabapentin 2 hours before anesthesia maintained with isoflurane had a MAC-sparing effect with no effect on hemodynamic variables or vital parameters of dogs.

Objective-To assess the feasibility of measuring cord dorsum potentials (CDPs) in anesthetized clinically normal dogs after caudal nerve stimulation, determine the intervertebral site of maximum amplitude and best waveform of the CDP, and evaluate the effects of neuromuscular blockade. Animals-8 male and 4 female dogs (age, 1 to 5 years). Procedures-Dogs were anesthetized, and CDPs were recorded via needles placed on the dorsal lamina at intervertebral spaces L1-2 through L7-S1. Caudal nerves were stimulated with monopolar electrodes inserted laterally to the level of the caudal vertebrae. Dogs were tested without and during neuromuscular blockade induced with atracurium besylate. The CDP latency and amplitude were determined from the largest amplitude tracings. Results-CDPs were recorded in 11 of 12 dogs without neuromuscular blockade and in all dogs during neuromuscular blockade. The CDP was largest and most isolated at the L4-5 intervertebral space (3 dogs) or the L5-6 intervertebral space (9 dogs); this site corresponded to the segment of insertion of the first caudal nerve. Onset latencies ranged from 2.0 to 4.7 milliseconds, and there was no effect of neuromuscular blockade on latencies. Amplitudes of the CDPs were highly variable for both experimental conditions. Conclusions and Clinical Relevance-CDPs were recorded from all dogs tested in the study; neuromuscular blockade was not critical for successful CDP recording but reduced muscle artifact. This technique may be useful as a tool to assess the caudal nerve roots in dogs suspected of having compressive lumbosacral disease or myelopathies affecting the lumbar intumescence.