Histological and finite element analysis of cell death due to irreversible electroporation.

Irreversible electroporation (IRE) has been shown to be an effective method of killing cells locally. In contrast to radiofrequency ablation, the mechanism by which cells are thought to die via IRE is the creation of pores in cell membranes, without substantial increase in tissue temperature. To determine the degree to which cell death is non-thermal, we evaluated IRE in porcine hepatocytes in vivo. Using pulse widths of 10 µs, bursts of 3 kV square-wave pulses were applied through a custom probe to the liver of an anesthetized pig. Affected tissue was evaluated histologically via stainings of hematoxylin & eosin (H&E), nitroblue tetrazolium (NBT) to monitor cell respiration and TUNEL to gauge apoptosis. Temperature was measured during the application of electroporation, and heat transfer was modeled via finite element analysis. Cell death was calculated via Arrhenius kinetics. Four distinct zones were observed within the ring return electrode; heat-fixed tissue, coagulation, necrotic, and viable. The Arrhenius damage integral estimated complete cell death only in the first zone, where the temperature exceeded 70°C, and partial or no cell death in the other zones, where maximum temperature was approximately 45°C. Except for a limited area near the electrode tip, cell death in IRE is predominantly due to a non-thermal mechanism.

Comparison of indirect and direct blood pressure monitoring in normotensive swine.

The gold standard for blood pressure measurement in pigs is direct monitoring of arterial pressure, but this is an invasive technique adding complexity to surgical procedures. We sought to compare direct measurements obtained via catheterization to more easily-obtained indirect measurements using a sphygmomanometer with an automated cuff. Simultaneous measurements via an arterial pressure transducer and a child-size cuff were performed in pigs undergoing abdominal surgical procedures under normotensive conditions. Correlation between direct and indirect measurements was good (r=0.881). Systolic blood pressures for the cuff were higher than those for arterial measurements, while diastolic pressures were lower for the cuff than arterial. A Bland-Altman analysis confirmed this bias at the extremes of the normotensive range. For highly accurate readings, especially under stressed conditions, direct arterial catheterization remains the preferred method of measuring blood pressure. When monitoring surgical procedures, the more convenient blood pressure cuff can provide reliable measurements.

The effect of measurement method on static weight distribution to all legs in dogs using the Quadruped Biofeedback System.

An application to measure static quadruped load distribution would be highly beneficial in the assessment of rehabilitation and lameness in many small animal patients. The scope of this study was to analyze the effects of confinement, location and local environment on the measurement of static quadruped load distribution as measured by the Quadruped Biofeedback System in normal dogs in order to better prescribe the use of this system. A prospective study of 20 healthy adult dogs was performed to evaluate effects on measurement on quadruped load distribution. Data collection in the form of mean load per extremity was recorded four times in five positions for a total of 20 measurements for each limb. A replicated cross-over design in which a mixed effect, repeated measures analysis of variance was used to test for main effects of treatment and end as well as their interaction. The effects of right vs. left within each end and for each end was analyzed for each treatment and significance of p < 0.01 was established. Measurements were taken from 20 healthy adult dogs with no obvious lameness at the time of data collection. Analysis by end suggests that measurements were affected in similar manners in comparable locations. Each method demonstrated consistency in measurement without any significant influence by day or session, suggesting that one standardized method be established for measurement. With standardization, the Quadruped Biofeedback System has potential use as a reliable instrument for the measurement of quadruped load distribution in dogs.

Tibial tuberosity advancement in 65 canine stifles.

The tibial tuberosity advancement (TTA) procedure was developed to treat dogs with cranial cruciate ligament deficient stifles. A retrospective, descriptive study was performed on 57 dogs that underwent unilateral or bilateral TTA. Medical records were reviewed and pre-, postoperative and follow-up radiographs were evaluated for patellar ligament-tibial plateau angle (alpha), distance of the tibial tuberosity advancement and progression of degenerative joint disease. A questionnaire was sent to all owners to obtain their assessment of the procedural outcome. Sixty-five stifles in 57 dogs received a TTA. Mean age was 5.2 +/- 2.5 years while mean weight was 39.7 +/- 11.9 kg. Eighteen breeds were represented with Labrador retrievers and mixed breeds predominating. The mean duration of lameness prior to surgery was 6.2 +/- 6.7 months, with a median lameness score of 3/4. Fifty-nine percent of cases encountered complications, the majority of which were minor. Major post-operative complications were uncommon but consisted of implant failure, tibial crest displacement and medial meniscal tears. The mean radiographic preoperative angle alpha was 100 degrees, while the postoperative was 95.5 degrees. Mean osteoarthrosis scores were significantly different between preoperative and follow-up radiographs with 67% of cases showing radiographic progression. Seventy percent of owners responded to the survey with overall outcome considered good to excellent in 90%. Activity level was improved in 90% of responses. TTA subjectively appears to be a useful alternative in the management of cranial cruciate ligament disease. Few severe complications were encountered. Good clinical outcome and owner satisfaction was reported with the procedure in this set of cases.

Surgical cytoreduction for the treatment of non-lymphoid vertebral and spinal cord neoplasms in cats: retrospective evaluation of 26 cases (1990-2005).

Medical records of 26 cats with non-lymphoid vertebral and spinal cord neoplasms treated surgically were reviewed to determine outcome and prognostic factors for survival. Of the factors examined, only tumour phenotype was significantly associated with survival. Osteosarcoma (3/26 cats) and meningioma (16/26 cats) were the most common malignant and benign tumours, respectively. The median survival time for cats with malignant neoplasms was 110.5 days, compared with 518 days for cats with benign tumours. Cytoreductive surgery resulted in clinical improvement in 25/26 cats, but local treatment failure occurred in 10/26 cats. Overall, 19/26 cats died of confirmed (12/19) or suspected (7/19) tumour-related causes, including all eight cats with malignant neoplasms. Results suggest that contemporary neurosurgical techniques commonly result in incomplete excision of feline non-lymphoid vertebral and spinal cord tumours but are efficacious at palliation of clinical signs of spinal cord dysfunction.

Evaluation of canine lumbosacral stenosis using intravenous contrast-enhanced computed tomography.

The objective of this study was to evaluate intravenous contrast-enhanced computed tomography as a technique for predicting the within-level location(s) of compressive soft tissues in the canine lumbosacral spine. Pre-operative intravenous contrast-enhanced computed tomography of the L5-S3 vertebral levels was performed in 12 consecutive large breed dogs with lumbosacral stenosis. The images were evaluated for enhancement of soft tissues by two radiologists who were unaware of the surgical findings. For each within-level location (dorsal canal, ventral canal, right lateral recess, left lateral recess) enhancement was classified as present, absent or equivocal. The results were compared with the results of surgical exploration and histopathology of excised tissues. The positive predictive values of intravenous contrast-enhanced computed tomography for compressive soft tissues involving the dorsal canal, ventral canal and lateral recesses were 83%, 100%, and 81% respectively. Negative predictive values for compressive soft tissues involving these locations were 29%, 50%, and 40% respectively.

Biomechanics of the thoracolumbar vertebral column of dogs during lateral bending.

OBJECTIVE: To study biomechanical characteristics of the normal and surgically altered canine thoracolumbar vertebral column to determine the effects of surgery and trauma on lateral stability. ANIMALS: The T13-L1 vertebral motion units of 48 mixed-breed dogs were dissected free of surrounding musculature and prepared for biomechanical testing by cross-pinning the vertebral bodies and mounting in polymethylmethacrylate. PROCEDURE: Normal and surgically altered spinal specimens were subjected to lateral bending. The mean slope of the bending moment versus angular displacement curve and the load to failure were compared between treatment groups and significance was determined by the method of least squares (P < 0.05). Specimens were surgically altered by facetectomy, lateral fenestration, diskectomy, and combinations of these procedures. Each specimen was subjected to lateral bending to failure at a rate of 2.5 cm/min in a swing arm bending jig designed to simulate 4-point bending and subject the specimen to pure bending. RESULTS: Only specimens undergoing diskectomy had a significant decrease in slope and load at failure. Unilateral and bilateral facetectomies and fenestration induced a nonsignificant decrease in stiffness, compared with control specimens. CONCLUSIONS: Fenestrations and facetectomies do not appear to increase the risk of injury to the canine thora-columbar spinal cord during lateral bending. CLINICAL RELEVANCE: Fenestrations and facetectomies, as used in routine laminectomies, may be performed without concern for significant destabilization of the spine in lateral bending; however, it is possible that thoracolumbar spinal fractures involving only the vertebral body may significantly destabilize the spine in all modes of bending.

Sacral fractures in dogs: a review of 32 cases.

Sacral fractures have not been described well in dogs. The records of 32 dogs diagnosed with sacral fractures were evaluated for neurologic deficits at presentation and discharge. Follow-up was in the form of telephone survey or physical examination at recheck. A score was assigned for each dog at presentation, discharge, and follow-up (0 for normal, 1 for minor deficits, and 2 for major deficits). Fractures located lateral to the sacral foramina were called abaxial, and those medial to the sacral foramina were called axial. Axial fractures had significantly more severe deficits at presentation (p = 0.00017) and discharge (p = 0.03063), but not at follow-up. Neurologic status did not improve significantly during hospitalization in either fracture group, but had improved significantly at follow-up.

Intracapsular repairs for cranial cruciate ligament ruptures.

Two intracapsular techniques and their modifications are described that can be regarded as the most accepted procedures in general use today. The difficulty in rationalizing a choice between intra- and extracapsular techniques is discussed with some principles being suggested to help the surgeon choose a technique.

Effect of left hepatic vein ligation on hepatic circulation, function, and microanatomy in dogs.

Eighteen healthy dogs were allotted to 3 groups (n = 6 dogs each). All dogs were evaluated at the beginning of the study by complete physical examination; total and differential WBC counts; serum biochemical analysis (alanine transaminase and alkaline phosphatase activities and bilirubin and albumin concentrations); sulfobromophthalein excretion, ammonia tolerance, and glucagon response testing; portal and intraparenchymal pressure determinations; operative mesenteric portography; and histologic assessment of hepatic biopsy specimens. The left hepatic vein was ligated completely in dogs of groups 1 and 2. Group-3 (control) dogs had a ligature placed loosely around the left hepatic vein. Dogs of groups 1 and 3 were reevaluated 24 hours after surgery by use of the aforementioned hematologic and biochemical tests. Group-1 dogs were reevaluated by use of portal and intraparenchymal pressure determinations, jejunal vein portography, and complete necropsy at 48 hours after surgery. At 4 weeks after surgery, dogs of groups 2 and 3 were reevaluated by use of all aforementioned tests. Results indicated transient hepatic congestion, which resolved by the fourth postoperative week. Longstanding effect on hepatic structure, circulation, or function was not found. We concluded that left hepatic vein ligation in clinically normal dogs does not cause severe or permanent liver damage.

Respiratory paralysis secondary to epidural anesthesia in a dog.

Transient respiratory paralysis developed in a 12-year-old spayed female Cocker Spaniel that received an anesthetic epidurally for excisional biopsy of perianal masses. Paralysis developed almost immediately after injection of 2.5 ml of 2% lidocaine (1 ml/4.9 kg), and was managed by mechanical ventilation and appropriate fluid therapy IV until spontaneous respiration returned. The respiratory paralysis was attributed to the excessive cranial extent of the anesthetic block.

Diskospondylitis in a kennel of dogs: clinicopathologic findings.

Selected aspects pertaining to the cause and pathogenesis of diskospondylitis were investigated in a kennel of 45 Airedale Terriers. Diskospondylitis was detected via spinal radiography in 17 male and 14 female dogs. Bacteria isolated from the coat, vagina, and urine of affected Airedales were not statistically different from those isolated from clinically normal Airedales. Serologic evidence of Brucella canis infection was not detected. There was no difference in response to thyrotropin stimulation tests between affected and clinically normal dogs. Necropsy findings in affected dogs included hypercellular renal glomeruli and pulmonary arterial thrombi.

Carpal hyperextension in two-month-old pups.

Abnormal carpal configuration was observed in a litter of ten 2-month-old pups raised in a research colony. Five of the pups were exercised daily, and carpal configuration returned to normal within 7 to 10 days. The other 5 pups remained confined, and their carpal configuration returned to normal after 6 weeks. It was concluded that poor muscle tone was responsible for the carpal hyperextension.

Ventral approach for stabilization of atlantoaxial subluxation secondary to odontoid fracture in a foal.

Atlantoaxial subluxation secondary to odontoid fracture in a 30-day-old foal was corrected by alignment of the atlantoaxial joint and stabilization with 2 ventrally placed dynamic compression plates. At 90 days after surgery, healing of the fracture, with adequate alignment of the atlantoaxial joint, was confirmed radiographically. The foal was only slightly tetraparetic at that time. At 1 year after surgery, the gait was normal. It was concluded that the technique has advantages over the use of Steinmann's pins or external coaptation for stabilization. The ventral approach allows decompression, anatomic alignment, and immediate stabilization of the subluxation. Potential complications of the ventral approach include laryngeal paralysis.

The effect of postoperative electromagnetic pulsing on canine posterior spinal fusions.

An experimental canine study was devised to evaluate the efficacy of a noninvasive adjunct to improve the rate and quality of the posterior fusion mass over the standard surgical technique. Ten large adult mongrel dogs underwent a three-level lumbar spinal fusion. Bone excised from the spinous processes was packed in removed facet joints and over the decorticated laminae. To insure rigid internal fixation, custom-made distraction instrumentation was placed bilaterally under the laminae of the vertebrae above and below the three fused vertebrae. Five dogs underwent electromagnetic pulsing, and five dogs acted as controls. Two dogs were sacrificed at 4, 6, 9, 12, and 15 weeks to assess the radiographic and histologic status of the fusion mass. Preoperative and preautopsy hematologic studies as well as gross and histologic autopsy specimens revealed no abnormalities attributable to the electromagnetic pulsing. High-resolution radiography and histologic studies showed earlier incorporation of the graft, improved new bone formation, and better organization of the fusion mass in the 4-, 6-, and 9-week stimulated specimens. However, by 12 and 15 weeks there did not appear to be any histologic or radiographic differences between the stimulated and control dogs. Although electromagnetic pulsing appears to produce an early accelerated osteogenic response, it does not appear to improve the overall results of primary canine spinal fusions.

Effect of localized trauma and temporary splinting on immature skeletal muscle and mobility of the femorotibial joint in the dog.

The right pelvic limbs of 2 groups of dogs were splinted in extension for 2 weeks. In 1 of these groups, the dogs were anesthetized and the distal portion of the quadriceps femoris muscle was traumatized before splinting. Two groups of control dogs were also used; neither group was splinted, but 1 group was anesthetized, and their muscles were traumatized. The range of motion of the femorotibial joint was measured in all 4 groups before splinting and 3 and 9 weeks after splinting. Types I and II muscle fiber diameters and percentages were also recorded at 3 and 9 weeks in all groups. Flexion of the femorotibial joint was limited after splinting (P less than 0.001), but less so after trauma and splinting than after splinting alone (P less than 0.0001). A reversible type I fiber atrophy occurred in most restricted muscles (P less than 0.001) and early type II fiber atrophy was seen in a few muscles after trauma and splinting (P less than 0.002). Multifocal fiber necrosis was the only irreversible change seen after 3 weeks of splinting with or without concurrent muscle trauma. Relative fiber percentages did not change appreciably during splinting or recovery.