Comparison of the neuromuscular blocking effects of cisatracurium during isoflurane or propofol anesthesia in dogs.

OBJECTIVE: To compare the neuromuscular blocking effects of cisatracurium during isoflurane versus propofol anesthesia in dogs. STUDY DESIGN: Prospective, randomized study. ANIMALS: A total of 20 healthy, client-owned dogs (16 females, four males) weighing 12.5-22 kg and aged 1-8 years. METHODS: Dogs undergoing elective surgery were randomized in equal numbers to an isoflurane (ISO) or propofol (PPF) group. Other drugs used during anesthesia were equal between groups. Single-twitch (ST) stimulation was used to monitor neuromuscular response. After recording the baseline ST (T0), cumulative doses of cisatracurium (0.05 mg kg-1) were administered intravenously until ST/T0 ≤5%. Effective doses 50 (ED50) and 95 (ED95) of cisatracurium in each group were calculated from group dose-response curves. Recovery of ST (TR) was defined as spontaneous recovery of ST to 80-120% of T0 remaining stable for 2 minutes. The ST after each dose of cisatracurium, duration 25% (time after the last dose until 25% recovery of TR), recovery index (time to recovery from 25% to 75% of TR) and duration to TR (time after the last dose until recovery of TR) were recorded. RESULTS: Incremental doses of cisatracurium, median (range), were 2 (1-3) in ISO and 4 (2-5) in PPF to achieve ≥95% depression of ST/T0 (p < 0.01). ED50 and ED95 were 20 µg kg-1 and 117 µg kg-1 in ISO and 128 µg kg-1 and 167 µg kg-1 in PPF, respectively. The duration 25%, recovery index and duration to TR, median (range), were longer in ISO [22.6 (10.3-24.3), 5.3 (3.0-7.8) and 36.1 (20.1-49.7) minutes, respectively] than in PPF [10.2 (6.8-16.5), 3.0 (2.0-3.8) and 17.7 (14.2-28.7) minutes, respectively] (p < 0.01). CONCLUSIONS AND CLINICAL RELEVANCE: Cisatracurium-induced neuromuscular blockade was significantly enhanced and prolonged by isoflurane compared with propofol.

Ultrasonographic features of insulinoma in six ferrets.

Insulinoma is a functional, insulin-secreting tumor, arising from the beta islet cells of the pancreas. It is one of the most common neoplasms in ferrets and has been associated with clinical signs of hypoglycemia, such as ptyalism, pawing at the mouth, seizures, lethargy, and coma. The ultrasonographic features of insulinoma in ferrets have not been previously reported. The purpose of this retrospective case series study was to describe the ultrasonographic features of confirmed insulinoma in a group of ferrets. Inclusion criteria were abdominal ultrasound examination and histological confirmed insulinoma by surgical biopsy. Six ferrets met the inclusion criteria, all of which had multiple hypoglycemic episodes. Ultrasonographic images were reviewed and the characteristics of the pancreatic nodules were recorded. Twenty-eight pancreatic nodules were observed in the six ferrets and were primarily hypoechoic (89.3%, 25/28) and homogenous (46.4%, 13/28) with a smooth margin (78.6%, 22/28). The distribution of the pancreatic nodules was 46.4% in the left lobe, 50% in the right lobe, and 3.6% in the body of the pancreas. The sizes of the pancreatic nodules varied from 1.5 × 1.5 to 4.1 × 5.6 mm. All of the pancreatic nodules removed from surgery were histopathologically confirmed as insulinoma. The findings indicated that insulinoma in ferrets could be detected through ultrasonography, which may facilitate diagnosis and preoperative surgical planning.

EFFECTS OF INTRAVENOUS DEXMEDETOMIDINE ON CARDIAC CHARACTERISTICS MEASURED USING RADIOGRAPHY AND ECHOCARDIOGRAPHY IN SIX HEALTHY DOGS.

Dexmedetomidine is a highly specific and selective α2-adrenergic receptor agonist widely used in dogs for sedation or analgesia. We hypothesized that dexmedetomidine may cause significant changes in radiographic and echocardiographic measurements. The objective of this prospective cross-sectional study was to test this hypothesis in a sample of six healthy dogs. Staff-owned dogs were recruited and received a single dose of dexmedetomidine 250 µg/m(2) intravenously. Thoracic radiography and echocardiography were performed 1 h before treatment, and repeated 10 and 30 min after treatment, respectively. One observer recorded cardiac measurements from radiographs and another observer recorded echocardiographic measurements. Vertebral heart score and cardiac size to thorax ratio on the ventrodorsal projection increased from 9.8 ± 0.6 v to 10.3 ± 0.7 v (P = 0.0007) and 0.61 ± 0.04 to 0.68 ± 0.03 (P = 0.0109), respectively. E point-to-septal separation and left ventricle internal diameter in diastole and systole increased from 2.4 ± 1.1 to 6.6 ± 1.9 mm, 32.3 ± 8.1 to 35.5 ± 8.8 mm, and 19.4 ± 6 to 27.0 ± 7.2 mm, respectively (P < 0.05). Fractional shortening and sphericity index decreased from 40.7 ± 5.8 to 24.4 ± 2.9%, and 1.81 ± 0.07 to 1.58 ± 0.04, respectively (P < 0.05). Moderate-to-severe mitral regurgitation and mild pulmonic regurgitation occurred in all dogs after dexmedetomidine administration. Findings indicated that dexmedetomidine could cause false-positive diagnoses of valvular regurgitation and cardiomegaly in dogs undergoing thoracic radiography and echocardiography.

A novel electroencephalographic evaluation of noxious stimulation during isoflurane anesthesia in dogs.

In veterinary clinical medicine, evaluating the balance between nociception and antinociception presents a great challenge for anesthesiologists during canine surgeries. Heart rate (HR) and mean arterial pressure (MAP) are suitable indexes for monitoring noxious stimuli during anesthesia. Frontal electroencephalography (EEG) records, including processed parameters, are recommended for evaluating nociceptive balance in anesthetized unconscious human patients, which is unexplored in veterinary medicine. Therefore, the objective is to explore the response of processed EEG parameters to noxious stimulation and elucidate the impact of noxious stimulation on frontal cortical activity in dogs anesthetized with 1.5% isoflurane. Fourteen dogs were included and underwent frontal EEG monitoring, measuring the patient state index (PSI) and spectral edge frequency (SEF) before and after administering noxious stimulation using the towel clamp method on the tail of each 1.5% isoflurane-anesthetized dog. As the noxious stimulation was applied, there was a simultaneous increase in PSI, HR, and MAP, with PSI exhibiting a drastic response. SEF, especially on the left side, also increased with noxious stimulation. In EEG power spectral analysis, the delta band was decreased, and the alpha and beta bands showed an increase following noxious stimulation, with a more profound elevation of beta bands on the left side. This study suggests that noxious stimulation brings asymmetric frontal cortical arousal, changing brain activity by suppressing delta waves and augmenting alpha and beta waves. Consequently, PSI seems to be a potential indicator for detecting stimuli in canine isoflurane anesthesia.

What Is Your Diagnosis?

In collaboration with the American College of Veterinary Radiology.

Effect of administering dexmedetomidine with or without atropine on cardiac troponin I level in isoflurane-anesthetized dogs.

We aimed to determine whether dexmedetomidine administration with or without atropine increases cardiac troponin I (cTnI) level in healthy dogs. We hypothesized that 10 µg/kg dexmedetomidine + atropine increases the cTnI level, whereas 5 µg/kg dexmedetomidine + atropine does not. Eighteen healthy, pet dogs that underwent an orthopedic surgery or ovariohysterectomy were included in this study. The dogs were randomly assigned to atropine (0.02 mg/kg)-dexmedetomidine (10 µg/kg), saline-dexmedetomidine (10 µg/kg), and atropine (0.02 mg/kg)-dexmedetomidine (5 µg/kg) groups. Each dog was premedicated with atropine or saline intramuscularly (IM). After 10 min, they were IM injected with dexmedetomidine (10 or 5 µg/kg)-morphine (0.5 mg/kg)-midazolam (0.2 mg/kg). Following this, anesthesia was induced after 10 min with propofol and maintained with isoflurane in 100% oxygen. The median plasma cTnI level at 6, 12 and 24 hr after premedication was significantly higher than that at baseline. The cTnI level in the atropine-dexmedetomidine (10 µg/kg) group was significantly higher than that in the saline-dexmedetomidine (10 µg/kg) and atropine-dexmedetomidine (5 µg/kg) groups at 6 and 12 hr after premedication. The cTnI level returned to normal within 72 hr after premedication in all groups. The administration of atropine in combination with 10 µg/kg dexmedetomidine increased the cTnI level, indicating subclinical myocardial damage.

A preliminary study comparing the sedative, cardiorespiratory, and histaminic-releasing effects of intramuscular and intravenous administration of pethidine (meperidine) with midazolam in healthy cats.

Pethidine is a synthetic opioid that is widely used in cats. However, the sedative, cardiorespiratory, and histaminic effects following administration of pethidine with midazolam in cats remain unclear. The objectives of this study were to evaluate and compare changes before and after intravenous (IV) and intramuscular (IM) administration of pethidine with midazolam in healthy cats. In this prospective randomized blind study, 12 cats were assigned equally to either the IV or IM treatment group. The IV group received pethidine 3 mg/kg and midazolam 0.1 mg/kg. The IM group received pethidine 6 mg/kg and midazolam 0.2 mg/kg. The sedative effects, heart rate, respiratory rate, non-invasive arterial blood pressures, and behavioral signs were recorded before and at 2, 5, 15, 30, 45, and 60 min after the injection. Blood samples were taken for an ELISA histamine assay at baseline and at 5 and 15 min after treatment. Cats that received IV treatment were rapidly induced a moderate degree of sedation but those received IM treatment were only mildly sedated. There was no significant difference in the cardiorespiratory values within and between the treatments over time. Plasma histamine concentrations increased by 3 and 5 times at 5 and 15 min after IV treatment, respectively, compared to baseline values. IM injections induced minimal changes in the plasma histamine concentration. In summary, intravenous pethidine with midazolam induced potentially superior sedative effects without serious side effects in clinically healthy cats. However, further studies with larger sample sizes are required to validate this finding.

Targeting l1 cell adhesion molecule using lentivirus-mediated short hairpin RNA interference reverses aggressiveness of oral squamous cell carcinoma.

The L1 cell adhesion molecule (L1CAM) has been implicated in tumor progression of many types of cancers, but its role in oral squamous cell carcinoma (OSCC) has not been investigated. In the present study, we demonstrated overexpression of L1CAM in OSCC cells, but not in normal keratinocytes, using both clinical specimens and cell lines. This overexpression demonstrated a strong correlation with less differentiation and a higher invasion potential of cancer cells, supporting the significance of L1CAM in human OSCC tumor progression. Targeting L1CAM gene expression in SCC4 cells overexpressing L1CAM using a lentivirus-mediated small hairpin RNA (shRNA) led to a significant reduction in cell proliferation in vitro via retardation of cell cycle at the G1 phase. In addition, shRNA knockdown of L1CAM strongly attenuated the migration and invasion of SCC4 cells, and this was also observed to parallel increased E-cadherin levels and decreased levels of vimentin, fibronectin, and Snail-family transcription factors, indicating that L1CAM expression was related to the epithelial-mesenchymal transition. Furthermore, while mice receiving orthotopically placed control SCC4 cells died within 40 days due to invasive tumor growth and regional lymph node metastasis, prolonged animal survival and complete suppression of tumor progression was observed in mice implanted with L1CAM-deficent SCC4 cells, further substantiating the fundamental importance of L1CAM in OSCC pathophysiology. Our findings suggested that L1CAM is a critical mediator of tumor progression in OSCC, and targeting L1CAM using lentivirus-mediated shRNA may be a useful molecular pharmaceutical approach for the treatment of advanced OSCC.

Functional expression of the recombinant ATPase of orf virus.

Nucleotide sequence analysis has indicated that the A32L gene of orf virus can encode an ATPase (Chan et al. in Gene 432:44-53, 2009). In this work, we cloned the A32L gene into a prokaryotic expression vector, and the recombinant protein was expressed and purified. The antigenicity of recombinant ATPase was examined by immunoblotting, and its identity was confirmed by mass spectrometry. The ATP hydrolysis function of the purified recombinant protein was examined, and our results showed that it exhibited the ATPase activity. Similar to other viral ATPases, the ATPase of orf virus remained active in the presence of different divalent ions; nevertheless, unlike other viral ATPases, our recombinant ATPase exhibited similar enzymatic activity in reaction buffers of different pH.

Comparison of the effects of morphine-lidocaine-ketamine and fentanyl-lidocaine-ketamine combinations administered as constant rate infusions on postprocedure rectal temperature in dogs.

OBJECTIVE: To compare the effects of morphine-lidocaine-ketamine (MLK) and fentanyl-lidocaine-ketamine (FLK) combinations administered as constant rate infusions (CRIs) during and after veterinary procedures on postprocedure rectal temperature in dogs. ANIMALS: 32 clinically normal client-owned dogs undergoing nonemergent procedures. PROCEDURES: Dogs were randomly assigned to receive an MLK or FLK combination (16 dogs/group). During the procedure, each dog received 2% lidocaine hydrochloride (1 mg/kg/h; both groups), ketamine hydrochloride (0.6 mg/kg/h; both groups), and morphine (0.36 mg/kg/h; MLK group) or fentanyl (4 µg/kg/h; FLK group) via CRI for analgesia; esophageal temperature was maintained at 37° to 39°C. At extubation, each drug dose in each assigned combination was halved and administered (via CRI) for 12 additional hours for postprocedure analgesia. Rectal temperature and other data were recorded at baseline (prior to administration of premedicants), extubation (0 hours), and 0.5, 1.5, 3, 6, and 12 hours thereafter. RESULTS: Mean postprocedure rectal temperature was significantly lower at each postextubation time point for the MLK group, compared with corresponding values for the FLK group. Compared with the baseline value, mean postprocedure rectal temperature was significantly lower at 0, 0.5, 1.5, and 3 hours for the FLK group and at all postprocedure time points for the MLK group. Hypothermia (rectal temperature < 37°C) was detected at ≥ 1 postprocedure time point more often in dogs in the MLK group (9/16) than in the FLK group (1/16). CONCLUSIONS AND CLINICAL RELEVANCE: Dogs that received an MLK combination for analgesia during and after a veterinary procedure developed hypothermia more commonly than did dogs that received an FLK combination under similar conditions.

Relationship between ultrasonographically determined renal dimensions and International Renal Interest Society stages in cats with chronic kidney disease.

BACKGROUND: The correlation between renal dimensions and renal function in cats with chronic kidney disease (CKD) is unclear. HYPOTHESIS/OBJECTIVES: To investigate the relationship between renal dimensions and CKD severity in cats using ultrasound examination. ANIMALS: Nineteen healthy cats and 30 cats with CKD. METHODS: Renal ultrasound images obtained between 2012 and 2016 were reviewed. Severity of CKD was determined using the International Renal Interest Society CKD staging system. Renal length, cortical thickness, medullary thickness, and corticomedullary ratio were measured, and the relationship between these renal dimensions and serum creatinine concentrations as well as differences in dimensions between the control and disease groups was investigated. The sensitivity and specificity of the renal dimensions for differentiation of the CKD also were evaluated. RESULTS: The disease group was subdivided into stage I to II (15 cats) and stage III to IV (15 cats) groups. Cortical thickness was significantly decreased in both disease groups and negatively correlated with disease severity. Compared with other renal dimensions, cortical thickness had a stronger linear correlation with the reciprocal of the serum creatinine concentration and superior diagnostic performance (Youden index: left kidney, 90.0% sensitivity and 94.7% specificity for a cutoff of 4.7 mm; right kidney, 83.3% sensitivity and 94.7% specificity for a cutoff of 4.5 mm). CONCLUSIONS AND CLINICAL IMPORTANCE: Decreased renal cortical thickness is observed in cats with loss of renal function. Measurement of cortical thickness using ultrasonography could be a useful method to evaluate the progression of CKD in cats.

Identification, expression and antigenic analysis of recombinant hemagglutinin proteins of canine distemper virus.

Canine distemper (CD) is a widely distributed disease of dogs, caused by the canine distemper virus (CDV). In the present study, the gene encoding the hemagglutinin (H) protein of a CDV isolate from central Taiwan was sequenced and compared with other strains. Sequence variations were noticed in the H gene from the field CDV strain that had previously been implicated in the increasing incidence of CD. To establish a serology-based diagnostic test, the full-length H protein, as well as five deletion mutants of a recombinant H protein of the local isolate, were produced using an E. coli expression system. Three truncated recombinant proteins with relatively high expression levels, designated HM3, HM4 and HM5, were used as antigens to examine their reactivity with canine sera. By using three negative sera and 17 CD-positive sera, the high specificity of recombinant H proteins was observed by ELISA. In addition, immunoblotting demonstrated that all three purified recombinant proteins exhibit an antigenic property recognized by the serum of a CD-suspected dog.

Clinical diagnosis and surgical management of diaphragmatic retroperitoneal perirenal fat and kidney herniation in a pet rabbit.

CASE DESCRIPTION A 6-year-old 2.08-kg (4.58-lb) neutered male Lionhead-mix pet rabbit (Oryctolagus cuniculus) was examined because of sneezing and increased respiratory effort. CLINICAL FINDINGS On the basis of the rabbit's radiographic findings, a diagnosis of diaphragmatic retroperitoneal perirenal fat and kidney herniation was made. Nine months later, physical examination revealed increased respiratory rate and effort and slightly decreased body weight. Thoracic radiography revealed decreased lung aeration and further craniomedial displacement of the right kidney, compared with the initial evaluation findings, suggesting progressive herniation of the retroperitoneal perirenal fat. TREATMENT AND OUTCOME During exploratory celiotomy, a tear in the right dorsal tendinous portion of the diaphragm was noted. The right kidney and perirenal fat were found to be displaced into the thorax. Diaphragmatic herniorrhaphy was performed after replacement of the right kidney and the perirenal fat in the retroperitoneal space. The rabbit recovered uneventfully from anesthesia and surgery. Clinical signs did not recur during the following 16 months. CLINICAL RELEVANCE For rabbits with increased respiratory effort, diaphragmatic retroperitoneal perirenal fat and kidney herniation should be included as a differential diagnosis. As illustrated by the case described in this report, appropriate surgical management can provide a successful outcome for affected pet rabbits.

Effects of cranberry extract on prevention of urinary tract infection in dogs and on adhesion of Escherichia coli to Madin-Darby canine kidney cells.

OBJECTIVE: To determine effects of cranberry extract on development of urinary tract infection (UTI) in dogs and on adherence of Escherichia coli to Madin-Darby canine kidney (MDCK) cells. ANIMALS: 12 client-owned dogs (in vivo experiment) and 6 client-owned dogs (in vitro experiment). PROCEDURES: 12 dogs with a history of recurrent UTI received an antimicrobial (n = 6) or cranberry extract (6) orally for 6 months. Dogs were monitored for a UTI. For the in vitro experiment, cranberry extract was orally administered to 6 dogs for 60 days. Voided urine samples were collected from each dog before and 30 and 60 days after onset of extract administration. Urine was evaluated by use of a bacteriostasis assay. An antiadhesion assay and microscopic examination were used to determine inhibition of bacterial adherence to MDCK cells. RESULTS: None of the 12 dogs developed a UTI. The bacteriostasis assay revealed no zone of inhibition for any urine samples. Bacterial adhesion was significantly reduced after culture with urine samples obtained at 30 and 60 days, compared with results for urine samples obtained before extract administration. Microscopic examination revealed that bacterial adherence to MDCK cells was significantly reduced after culture with urine samples obtained at 30 and 60 days, compared with results after culture with urine samples obtained before extract administration. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of cranberry extract prevented development of a UTI and prevented E coli adherence to MDCK cells, which may indicate it has benefit for preventing UTIs in dogs.

Emphysematous pyometra secondary to Enterococcus avium infection in a dog.

A 5-year-old female intact Mastiff dog was presented with a history of vaginal discharge for 1 day. Physical examination revealed a sanguineo-purulent vaginal discharge and systemic inflammatory response syndrome. Abdominal radiographs showed several dilated and gas- filled tubular loops. The differential diagnoses included emphysematous pyometra or small intestinal mechanical ileus. Surgical exploration of the abdomen demonstrated a severely dilated and gas-filled uterus, and emphysematous pyometra was confirmed. The patient's clinical signs resolved after ovariohysterectomy. Histopathology revealed mild endometrial cystic hyperplasia with infiltration of inflammatory cells in the superficial endometrial epithelia. Enterococcus avium, an α-hemolytic gram-positive coccus, was isolated from the uterus. This paper highlights the radiographic features of emphysematous pyometra and a pathogen that has never been reported to be associated with canine pyometra previously.

On the TATA box and transcriptional start site of gE gene of pseudorabies virus: a comparison of three methods.

Using the upstream region of glycoprotein E gene of pseudorabies virus (PRV) as a model, a method based on reverse transcriptase-polymerase chain reaction (RT-PCR) for the determination of transcriptional start site was developed. The conventional primer extension method was used to determine the start site. Comparing the results obtained by these two methods, a good agreement on the location of start site was achieved. In addition, a computer program was used to predict the transcriptional start site, and the predicted site was found to be close to the two sites obtained by experiments. Based on the transcriptional start site data and on the general knowledge of the eukaryotic gene, the TATA box of gE gene was assigned at approximately 33 bp upstream to the start site.

Lytic infection of pseudorabies virus in the presence of spermine, spermidine, or DFMO.

The effects of polyamines (spermine or spermidine) and DFMO (an ornithine decarboxylase inhibitor) on the infection of LM (tk-) cells by pseudorabies virus (PRV) were investigated. Results from radioactive methionine labeling showed that the synthesis of viral proteins was not affected; however, the expression of a distinctive cellular protein ( approximately 27 kDa) was induced after the treatment of spermine or spermidine. Using plaque assay, we found that the plaque formation of PRV was not affected by these three reagents either. Furthermore, the effects of these drugs on the transcription of PRV immediate-early gene (IE) promoter were examined by CAT assay, and results showed weak stimulation of transcription by these drugs. Taken together, our results demonstrated that lytic infection of PRV was not influenced by addition of exogenous polyamines or depletion of endogenous polyamines; this conclusion was similar to earlier studies by using herpes simplex virus.

Examination of meat components in commercial dog and cat feed by using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) technique.

It has been shown that certain slow neurological diseases such as bovine spongiform encephalopathy (also known as "mad cow" disease) could be transmitted through contaminated food intake by animals; therefore, the examination of meat components in commercial feeds is important for the control of the disease in public health. The combination of polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) technique was applied to examine the meat components in dog and cat commercial feeds. The partial nucleotide sequence (359 bp) of animal mitochondrial cytochrome b (cytb, CYT) gene was amplified by PCR and then digested with restriction enzyme Alu I or Mbo I. In this work, eight brands of commercial dog and cat feeds available in Taiwan were examined. All brands of dog feeds that were tested contained meat from four different animals (cattle, pig, goat and chicken). In cat feeds, the chicken meat was found in five out of eight brands.

Decreased postoperative C-reactive protein production in dogs with pyometra through the use of low-dose ketamine.

OBJECTIVE: To determine the effects of subanesthetic ketamine in dogs with pyometra on C-reactive protein (CRP) concentrations following surgery. DESIGN: Prospective, nonconcealed, alternating allocation controlled trial. SETTING: Veterinary teaching hospital. ANIMALS: Sixteen dogs diagnosed with pyometra. INTERVENTIONS: The tentative diagnosis of canine pyometra was based on compatible history, physical examination findings, ultrasonographic findings, and hematological evaluation. Two different anesthesia and analgesic protocols with and without low-dose ketamine were used during and following ovariohysterectomy in 16 female dogs (n = 8 per group) that were diagnosed with naturally occurring pyometra. Dogs were sequentially allocated to treatment groups in an alternating fashion without concealment. Serum was collected before, 24, and 48 hours after surgery for CRP measurement. MEASUREMENTS AND MAIN RESULTS: Perioperative physical parameters in the 2 groups of dogs were similar. The serum concentrations of CRP in both groups were essentially the same before surgery, but significantly increased in the control group and decreased in ketamine group at 48 hours after surgery. CONCLUSIONS: Low-dose ketamine attenuated the postoperative concentration of serum CRP in dogs with pyometra compared with dogs that did not receive ketamine in the perioperative period. Further studies are warranted to determine the clinical implications of these findings.