Evaluation of injectate distribution after ultrasound-guided peribulbar injections in canine cadavers.

OBJECTIVE: To describe a contrast medium distribution pattern after peribulbar injection in cadavers. STUDY DESIGN: A cadaveric study. ANIMALS: A group of eight male dogs and four canine cadaver heads. METHODS: In phase 1, ultrasound-guided peribulbar injections (1.0 mL dye) were performed in four orbits using a subzygomatic approach. After dissection, the procedure was considered successful if 1) no dye was found in the retrobulbar space, 2) the peribulbar compartment was stained and 3) the dye reached the orbital fissure. In phase 2, using 16 orbits, the same technique was used to inject 0.1 mL kg-1 contrast medium into (group A, n = 8) or outside (group B, n = 8) the peribulbar compartment. After freezing at -20°C, orbits were imaged using computed tomography, and the contrast medium distribution was analysed. RESULTS: In phase 1, the procedure was successful in all four eyes. In phase 2, two orbits (one in each group) were removed from the study: one was not injected and the other was injected twice. The contrast medium was found inside the peribulbar compartment in six of seven (85.7%) group A orbits and two of seven (28.6%) group B orbits. In all of these cases, distribution to the orbital fissure, rostral alar foramen, mandibular foramen and inside the skull was observed. The oval foramen was reached in three of seven (42.9%) group A orbits and one of seven (14.3%) group B orbits. In one/seven (14.3%) group A orbits and five/seven (71.4%) group B orbits, the contrast medium was found inside the masseter and temporal muscles, and no distribution towards the aforementioned structures was observed. CONCLUSIONS AND CLINICAL RELEVANCE: Ultrasound-guided peribulbar injection of 0.1 mL kg-1 contrast medium in canine cadavers can lead to an extensive distribution of the injectate that reaches the orbital fissure, the rostral alar, oval, and mandibular foramina and can be found intra-cranially.

Anatomical and ultrasonographic study of the femoral nerve within the iliopsoas muscle in beagle dogs and cats.

OBJECTIVES: An ultrasound (US)-guided ventral suprainguinal approach to block the femoral nerve (FN) within the iliopsoas muscle (IPM) has recently been described in dogs. The goal of the present study was to provide the operator with additional information to locate the FN within the IPM in dogs and cats using US. STUDY DESIGN: The study was carried out in three phases: a dissection of the FN (phase 1); an in vivo US-assisted nerve study (phase 2), and an anatomical cross-sectional study (phase 3). ANIMALS: Nine healthy adult beagle dogs and nine healthy adult cats. METHODS: Dissections were performed to investigate the anatomical characteristics of the FN and its related structures in one dog and one cat. Ultrasound scans of the left and right FN were performed in eight dogs and eight cats. The FN diameter and the distance between the FN and the external iliac artery (EIA) in US images and in anatomical cryosections were measured. RESULTS: The median FN diameter did not differ significantly between cats and dogs (1.1 mm versus 1.0 mm) or between the two techniques (US versus anatomical cross-sectional study) (1.1 mm versus 1.1 mm in dogs; 1.0 mm versus 1.1 mm in cats). The US and anatomical measurements of the median distances between the FN and EIA differed significantly between dogs and cats (8.2 mm versus 5.8 mm by US; 5.7 mm versus 4.8 mm in the anatomical study). CONCLUSIONS AND CLINICAL RELEVANCE: The distance between the EIA and FN is reproducible in beagle dogs and cats and can be used in locating the FN within the IPM.

Ultrasound guidance to approach the femoral nerve in the iliopsoas muscle: a preliminary study in the dog.

OBJECTIVE: To describe the location of the femoral nerve within the iliopsoas muscle and determine the feasibility and potential complications of an ultrasound-guided block. STUDY DESIGN: Prospective experimental trial. ANIMALS: Sixteen adult dogs, median weight 14.3 (range 3-37) kg. METHODS: Phase I. Computed tomographic images of the iliopsoas muscle and femoral nerve were analyzed in two dogs. Phase II. The location of the femoral nerve within the iliopsoas muscle was determined via ultrasonography in 11 healthy anaesthetized dogs. Phase III. Peripheral nerve stimulation and ultrasound were combined to perform femoral nerve blocks in three dogs. RESULTS: Using computed tomography, the femoral nerve and its L4-L5 roots were identified within the iliopsoas muscle. The nerve itself was traced until it branched off into the quadriceps femoris muscle. Using ultrasonography, it was possible to observe the femoral nerve in nine dogs (82%). Starting at the dorsal third of the iliopsoas muscle, its path was traced in a caudo-ventro-lateral direction, emerging from the iliopsoas muscle shortly before passing through the muscular lacuna where it became very difficult to identify. An ultrasound-guided femoral nerve approach was carried out successfully in all three dogs. CONCLUSIONS: It is possible to approach the femoral nerve using combined ultrasound guidance and peripheral nerve stimulation to a closer proximal point than previously described. CLINICAL RELEVANCE: The ultrasound-guided approach of the femoral nerve within the iliopsoas muscle has the potential to become an additional approach.

Hypothermic storage and cryopreservation of hepatocytes: the protective effect of alginate gel against cell damages.

Hepatocyte-based therapy has been proposed as an alternative to organ transplantation in the treatment of liver disorders. In the clinical context, a major issue is the constant supply of quality assurance-controlled hepatocytes, thereby requiring their cold storage in good conditions. We have analyzed the protective effects of alginate entrapment of rat hepatocytes after either 24 or 48 h of hypothermic storage or cryopreservation on the cell viability, cell yield, both mitochondrial and other cytoplasmic functional activities, and apoptosis. Decrease in viability, as evaluated by the MTT inclusion test, was 4% and 13% (24 h at 4 degrees C), 15% and 33% (48 h at 4 degrees C), and 9% and 19% (liquid nitrogen) for entrapped and free suspended hepatocytes, respectively. Viable cell yields were 86 +/- 8% and 51 +/- 6% for cryopreserved entrapped and free suspended hepatocytes, respectively. The mitochondrial (MTS assay), 7-ethoxyresorufin O-deethylase (EROD), and glutathione-S-transferase (GST) activities were better preserved in entrapped than in free suspended hepatocytes. Both hypothermic storage and cryopreservation were found to induce early caspase-3-like activities, being always much lower in entrapped hepatocytes, particularly after cryopreservation (98.4 +/- 42.4 vs. 6.4 +/- 4.0 fluorescence arbitrary units/hours/microg protein). Thus, cold-induced apoptosis in hepatocytes can be significantly reduced following their entrapment within alginate gel beads and this is associated with an improvement of both their viability and function.

Method for monitoring alginate released in biological fluids by high-performance anion-exchange chromatography with pulsed amperometric detection.

The use of alginate-entrapped cells in cell therapy requires a method for monitoring possible released compound within biological fluids following either their implantation or inoculation in artificial organs. Oligomannuronic and oligoguluronic acids were prepared by enzymatic depolymerization with alginate lyase from Pseudomonas alginovora, characterized by high-performance anion-exchange chromatography with pulsed amperometric detection and quantitated in human, pig and rabbit blood, urine and tissue samples. The method was tested for linearity and detection limit, accuracy, intra- and inter-day precision. The limit of detection was 3 microgram/ml in both urine and plasma and 5 mg/g of tissues. The relative standard deviations (RSDs) of intra-day precision were 6.0-16.6% and 4.8-8.7% in plasma and urine, respectively; the RSDs of inter-day precision were 5.1-14.4% and 5.0-11.6% in plasma and urine, respectively. Thus, this method appears suitable for the measurement of released alginate from entrapped cells used in cell therapy.

Anatomical and experimental studies of brachial plexus, sciatic, and femoral nerve-location using peripheral nerve stimulation in the dog.

OBJECTIVE: To investigate the anatomy of the brachial plexus, sciatic, and femoral nerves for the use of a peripheral nerve-stimulator to perform nerve blocks in dogs. STUDY DESIGN: Prospective experimental trial. ANIMALS: Four canine cadavers and four healthy adult dogs weighing 23 +/- 2.5 kg. METHODS: Phase I: in four canine cadavers, an anatomical study was conducted to evaluate accurate needle-insertion techniques. Phase II: the utility of these techniques, and the value of electrostimulation, were evaluated in four anesthetized dogs in lateral recumbency (medetomidine, 5 microg kg(-1)/ketamine 5 mg kg(-1)) using an electrical stimulator and shielded needles. RESULTS: For the brachial plexus, the needle was inserted cranial to the acromion, medial to the subscapularis muscle, at an angle of approximately 20-30 degrees in relation to a plane vertical to the surface on which the animal was lying, oriented parallel to the long axis of the animal, in a ventro-caudal direction. For the sciatic nerve, the needle was inserted just cranial to the sacrotuberous ligament, through the gluteus superficialis muscle, at an angle of approximately 60 degrees in relation to the horizontal plane, in a ventro-cranial direction, and up to the level of the ischium. For the femoral nerve, the needle was inserted perpendicular to the skin, just cranial to the femoral artery, and directed a little caudally. Using a peripheral nerve-stimulator, all nerves were located, and muscle contractions were elicited at a current of 0.2-0.4 mA. No complications were observed during the procedures. CONCLUSION: Electrostimulation of peripheral nerves is useful in locating the branches of the brachial plexus as well as the sciatic and femoral nerves in dogs. CLINICAL RELEVANCE: Peripheral nerve stimulation increases the reliability of a nerve block when compared with blind needle-insertion.

Electrical nerve stimulation to facilitate placement of an indwelling catheter for repeated brachial plexus block in a traumatized dog.

A 10-year-old, 7 kg, male mixed breed dog was presented, following a dog fight, with open fractures of the radius and ulna and extensive skin and muscle lesions. Using electric nerve stimulation, an indwelling catheter was positioned in the axillary space and 2 mg kg(-1) bupivacaine was injected. Complete sensory and motor blocks were obtained allowing amputation distal to the non-traumatized tissue and debridement of the limb. The injection of bupivicaine was repeated 5 hours after the first bolus and a similar result was obtained in term of muscle relaxation and pain alleviation. Subsequent injections were not successful in providing the same effect, most likely as a result of catheter displacement and alternate means of analgesia were used.

Preservation of the fistula for reconstruction of the anal canal and the anus in atresia ani and rectovestibular fistula in 2 dogs.

OBJECTIVE: To report a technique for correction of atresia ani and rectovestibular fistula with fistula preservation, and outcome in 2 dogs. STUDY DESIGN: Case report. ANIMALS: Two intact female puppies. METHODS: The fistula was approached by episiotomy. A vertical median perineal incision was made starting dorsal to the anal region and extended to surround the fistula. The rectum and vagina were separated, and the end of the fistula was sutured to the skin. RESULTS: Early postoperative complications included constipation, fecal incontinence, and perineal soiling, but long-term outcome was good in both dogs. CONCLUSION: With careful dissection, the fistula and internal anal sphincter can be preserved and used in the surgical reconstruction of the anal canal and anus. CLINICAL RELEVANCE: Preservation of the fistula may provide an internal sphincter that contributes to a better functional result during correction of atresia ani. A surgical approach that combines episiotomy and perineal incision for separation of the vagina and rectum, and preserves the fistula may decrease the risk of dehiscence.

Reduced encephalopathy in pigs with ischemia-induced acute hepatic failure treated with a bioartificial liver containing alginate-entrapped hepatocytes.

OBJECTIVE: To analyze the effects of an extracorporeal bioartificial liver containing alginate bead-entrapped hepatocytes on pigs with ischemia-induced acute hepatic failure. DESIGN: Prospective animal study. SETTING: University and INSERM laboratory. SUBJECTS: Fifteen Large White/Pietrin female pigs weighing 20-30 kg. INTERVENTIONS: Acute hepatic failure was induced by end-to-side portocaval shunt and ligature of the whole porta hepatitis. The bioartificial liver was in a thermostabilized column, containing a fluidized bed of alginate beads that embedded porcine hepatocytes, connected to a plasmapheresis system. Intracranial pressure; survival; ammonia, total bilirubin, aminotransferases, alkaline phosphatase, and lactate concentrations; and clotting factors were studied. The groups were pigs with acute hepatic failure (group 1, n = 4), pigs with acute hepatic failure treated with bioartificial liver containing empty beads (group 2, n = 4), or porcine hepatocytes (group 3, n = 5). MEASUREMENTS AND MAIN RESULTS: In group 1, survival of pigs averaged 10.9 +/- 1.0 hrs; intracranial pressure reached 32.3 +/- 3.8 mm Hg and was associated with coma and cerebral edema. After connection to the bioartificial liver, the survival of acute hepatic failure pigs was 12.1 +/- 1.4 hrs in group 2 and 14.8 +/- 2.5 hrs in group 3. In group 3, intracranial pressure and bilirubin concentrations were reduced significantly compared with both group 1 and group 2. Neither signs of encephalopathy nor cerebral edema was observed in any animal of group 3. In all animals, plasma ammonium, aminotransferases, alkaline phosphatase, and lactate concentrations increased and clotting factors decreased with no significant differences between the three groups. Autopsy revealed a total necrosis of the liver, which was histologically confirmed. CONCLUSIONS: The ischemia-induced model of acute hepatic failure in pigs is reproducible and provides measurable clinical and biological features. A bioartificial liver containing alginate bead-entrapped hepatocytes improves the signs of encephalopathy in pigs with ischemia-induced acute hepatic failure, suggesting that the bioartificial liver can clear out toxic compounds that are released from necrotic livers.

Follow-up by one- and two-dimensional NMR of plasma from pigs with ischemia-induced acute liver failure treated with a bioartificial liver.

Hepatic encephalopathy may occur following acute hepatic failure (AHF), which results in the release of toxic compounds from the injured liver. These compounds, which induce cerebral edema, are not well characterized, yet. The aim of this study was to evaluate the potential interest of NMR spectroscopy in the follow-up of different plasma compounds in pigs with ischemia-induced fulminant hepatic failure treated or not with a bioartificial liver (BAL), which has been previously shown to improve the neurological status of the animals. Qualitative analysis of pig plasma was achieved by one-dimensional-(1)H CPMG, two-dimensional homonuclear (1)H-(1)H TOCSY CPMG and heteronuclear (1)H-(13)C HSQC sequences. Semi-quantitative analysis of selected plasma metabolites along the disease evolution was carried out on pigs with ischemia-induced AHF treated with the BAL containing alginate beads with or without hepatocytes. A quantitative longitudinal follow-up was performed on characteristic metabolites via a one-dimensional CPMG sequence, including choline, glutamine, N-acetyl-glucosamine (NAG), pyruvate and trimethylamine-N-oxide (TMAO). The concentrations of choline and TMAO increased from the beginning to the end in animals treated with the BAL containing alginate beads without hepatocytes. Treatment of pigs with BAL containing hepatocytes resulted in an improvement of survival, the plasma concentrations of choline and TMAO being decreased in three out of five animals. Thus, NMR spectroscopy is a useful approach for the identification of toxic compounds which are involved in hepatic encephalopathy associated with AHF. These compounds can be cleared by a BAL resulting in the improvement of survival and neurological parameters of the animals.

Anatomical and experimental studies of brachial plexus, sciatic, and femoral nerve-location using peripheral nerve stimulation in the dog.

OBJECTIVE: To investigate the anatomy of the brachial plexus, sciatic, and femoral nerves for the use of a peripheral nerve-stimulator to perform nerve blocks in dogs. STUDY DESIGN: Prospective experimental trial. ANIMALS: Four canine cadavers and four healthy adult dogs weighing 23 +/- 2.5 kg. METHODS: Phase I: in four canine cadavers, an anatomical study was conducted to evaluate accurate needle-insertion techniques. Phase II: the utility of these techniques, and the value of electrostimulation, were evaluated in four anesthetized dogs in lateral recumbency (medetomidine, 5 microg kg(-1)/ketamine 5 mg kg(-1)) using an electrical stimulator and shielded needles. RESULTS: For the brachial plexus, the needle was inserted cranial to the acromion, medial to the subscapularis muscle, at an angle of approximately 20-30 degrees in relation to a plane vertical to the surface on which the animal was lying, oriented parallel to the long axis of the animal, in a ventro-caudal direction. For the sciatic nerve, the needle was inserted just cranial to the sacrotuberous ligament, through the gluteus superficialis muscle, at an angle of approximately 60 degrees in relation to the horizontal plane, in a ventro-cranial direction, and up to the level of the ischium. For the femoral nerve, the needle was inserted perpendicular to the skin, just cranial to the femoral artery, and directed a little caudally. Using a peripheral nerve-stimulator, all nerves were located, and muscle contractions were elicited at a current of 0.2-0.4 mA. No complications were observed during the procedures. CONCLUSION: Electrostimulation of peripheral nerves is useful in locating the branches of the brachial plexus as well as the sciatic and femoral nerves in dogs. CLINICAL RELEVANCE: Peripheral nerve stimulation increases the reliability of a nerve block when compared with blind needle-insertion.