Evaluation of immunologic parameters in canine glioma patients treated with an oncolytic herpes virus.

Aim: To molecularly characterize the tumor microenvironment and evaluate immunologic parameters in canine glioma patients before and after treatment with oncolytic human IL-12-expressing herpes simplex virus (M032) and in treatment naïve canine gliomas. Methods: We assessed pet dogs with sporadically occurring gliomas enrolled in Stage 1 of a veterinary clinical trial that was designed to establish the safety of intratumoral oncoviral therapy with M032, a genetically modified oncolytic herpes simplex virus. Specimens from dogs in the trial and dogs not enrolled in the trial were evaluated with immunohistochemistry, NanoString, Luminex cytokine profiling, and multi-parameter flow cytometry. Results: Treatment-naive canine glioma microenvironment had enrichment of Iba1 positive macrophages and minimal numbers of T and B cells, consistent with previous studies identifying these tumors as immunologically "cold". NanoString mRNA profiling revealed enrichment for tumor intrinsic pathways consistent with suppression of tumor-specific immunity and support of tumor progression. Oncolytic viral treatment induced an intratumoral mRNA transcription signature of tumor-specific immune responses in 83% (5/6) of canine glioma patients. Changes included mRNA signatures corresponding with interferon signaling, lymphoid and myeloid cell activation, recruitment, and T and B cell immunity. Multiplexed protein analysis identified a subset of oligodendroglioma subjects with increased concentrations of IL-2, IL-7, IL-6, IL-10, IL-15, TNFα, GM-CSF between 14 and 28 days after treatment, with evidence of CD4+ T cell activation and modulation of IL-4 and IFNγ production in CD4+ and CD8+ T cells isolated from peripheral blood. Conclusion: These findings indicate that M032 modulates the tumor-immune microenvironment in the canine glioma model.

Central nervous system lymphoma in 18 dogs (2001 to 2015).

OBJECTIVES: To investigate the outcome of dogs with central nervous system lymphoma. MATERIALS AND METHODS: A multi-center, retrospective, observational study was conducted by reviewing medical records of 18 cases of central nervous system lymphoma from seven institutions. RESULTS: Diagnosis of lymphoma was made through cerebrospinal fluid analysis, histopathology, flow cytometry of the cerebrospinal fluid, and cytology of cerebrospinal fluid, lymph node or spleen with correlated imaging. A total of 15 of 18 dogs received specific treatment other than prednisone. Three dogs underwent chemotherapy and radiation therapy after surgical decompression, five dogs underwent chemotherapy, two dogs underwent radiation therapy after surgical decompression, three dogs underwent chemotherapy after surgical decompression and two dogs underwent radiation therapy and chemotherapy. Only one dog received prednisone, and two dogs did not receive any treatment. Overall, the median survival time was 171 days (range 1 to 1942 days). CLINICAL SIGNIFICANCE: Dogs receiving any type of treatment for central nervous system lymphoma lived longer than cases described in previous historical reports. Further studies are needed to elucidate the importance of specific treatment modalities.

The effect of electromagnetic pulsing on posterior lumbar spinal fusions in dogs.

This study evaluated the effect of pulsed electromagnetic fields (PEMF) on the healing of lumbar spinal fusions. Bilateral posterior facet fusions were performed at L1-2 and L4-5 in 24 adult mongrel dogs. After surgery, eight animals were stimulated with a pulse burst type signal (PEMF) for 30 minutes a day, and eight animals were stimulated with the same PEMF for 60 minutes a day. The remaining eight animals received no active PEMF stimulation and served as controls. Four animals from each group were euthanatized at 6 and 12 weeks, and the facet fusions were evaluated using high resolution radiographs and routine histology. No statistical difference in the radiographic or histologic appearance of the fusion mass could be detected between the stimulated and control groups at either 6 or 12 weeks. The results of this study suggest that PEMF stimulation had no effect on the healing of the primary posterior spinal fusions in this controlled experimental canine model.

Idiopathic polyneuropathy in Alaskan malamutes.

Clinical and morphologic features of a progressive polyneuropathy in young mature Alaskan Malamutes are described. Clinical signs included progressive paraparesis, synchronous pelvic limb gait, exercise intolerance, hyperesthesia, hyporeflexia, muscle atrophy, and tetraplegia. Electromyographic testing revealed diffuse fibrillation potentials and positive sharp waves in limb muscles, especially in muscles below the elbow and stifle. Pathologic findings in skeletal muscles and peripheral nerves included neurogenic muscle atrophy, focal or diffuse loss of myelinated nerve fibers, myelinoaxonal necrosis, and variable demyelination or remyelination. Ultrastructural changes included axonal degeneration, presence of numerous Büngner bands, and denervated Schwann cell subunits. The nature and distribution of abnormal electrophysiologic and pathologic findings were suggestive of a distal sensorimotor polyneuropathy, which we have termed idiopathic polyneuropathy of Alaskan Malamutes to distinguish this condition from hereditary polyneuropathy of Norwegian Alaskan Malamutes, last described in 1982.

Spinal-evoked potentials in dogs with acute compressive thoracolumbar spinal cord disease.

Spinal evoked potentials (SpEP) were recorded on an electromyograph from electrodes placed percutaneously in the ligamentum flava at the lumbosacral junction and between the 10th and 11th thoracic vertebrae following tibial nerve stimulation in 31 anesthetized dogs with acute compressive spinal cord injuries. The neurologic status of each dog was determined by clinical examination before SpEP recordings, and the neurologic status was monitored for 2 months in dogs that had surgical or conservative treatment. Two months after spinal injury, the response to treatment (outcome) of each dog was evaluated and graded as favorable (ambulatory and urinary continent) or unfavorable (nonambulatory, urinary incontinent, or euthanatized with confirmation of myelomalacia). Onset latencies, conduction velocities, amplitudes and durations of the wave forms, and the ratio of conduction velocity to combined durations of the first positive (P1) and first negative (N1) waves (CV/DPN index) were determined and were compared with reference data from clinically normal (control) dogs. Single SpEP recordings were of value in determining the prognosis for recovery. Significant differences were not found in the L7-S1 recordings between the reference (control) and spinal injury groups. Analysis of data from the T10-11 recordings indicated significant differences between the reference and spinal injury groups and between the favorable and unfavorable outcome groups within the spinal injury group. A CV/DPN index was less than 30 in dogs with unfavorable outcomes and greater than 30 in dogs with favorable outcomes. Stepwise discriminant analysis of data from the spinal injury group predicted outcome correctly in all dogs.

Combined Kirschner-Ehmer apparatus and dorsal spinal plate fixation of caudal lumbar fractures in dogs: biomechanical properties.

The strength and rigidity of a new surgical technique for the stabilization of caudal lumbar fractures in dogs, using a Kirschner-Ehmer apparatus and a dorsal spinal plate (KE/DSP), were compared with 2 other methods of internal spinal fixation and with intact (control) spines, using a spinal test system that subjected the spines to 4-point bending. The fixation devices were applied to isolated canine lumbosacral spines (L1 to S3) from cadavers. A complete spinal separation was made in the spine implant specimens at L5-L6 by sharp dissection of all ligamentous structures connecting the two vertebrae. Bending moment vs L5-L6 angular deformation curves, and rigidity and load sustained at 10 degrees angular deformation (failure) were recorded for each fixation method and for the control spines. Values were compared by statistical analysis. The combined KE/DSP fixation and a combined vertebral body plate/dorsal spinal plate (VBP/DSP) fixation were stronger and more rigid than were the control spines and those fixed with a modified segmental-fixation method (P less than 0.05). There were no statistical differences in strength and rigidity between the 2 combined-fixation techniques. Although the VBP/DSP technique is not applicable to clinical caudal lumbar (L5-L6) fractures, it was compared in this study to the KE/DSP technique because a similar VBP/DSP technique was reported strongest in a similar study of L3-L4 simulated fractures, compared with 3 other spinal-fixation techniques that have been used in repair of caudal lumbar fractures. The technique has been used successfully in 6 dogs with caudal lumbar fractures.

Laryngeal paralysis-polyneuropathy complex in young Dalmatians.

Clinical, morphologic, and morphometric findings are reported in 14 young Dalmatians with laryngeal paralysis. Neurologic signs, including megaesophagus, were observed in 13 of 14 dogs. Electromyographic abnormalities included fibrillation potentials and positive sharp waves in laryngeal, esophageal, facial, and distal appendicular muscles. Neurogenic atrophy was detected in intrinsic laryngeal and appendicular skeletal muscles. A diffuse, generalized polyneuropathy, dominated by axonal degeneration, was observed in recurrent laryngeal and appendicular peripheral nerves. Results of quantitative studies, using single teased fiber and cross-sectional nerve preparations, indicated that changes were more severe in distal parts of peripheral nerves, with preferential loss of medium sized (5.5 to 8 microns) and large-caliber (8.5 to 12 microns) myelinated nerve fibers. Ultrastructural alterations were observed in myelinated and unmyelinated nerve fibers. The term laryngeal paralysis-polyneuropathy complex is proposed for this apparent dying-back disorder, which is clinically, electrophysiologically, and pathologically different from laryngeal paralysis in young Bouvier des Flandres and Siberian Huskies. Prognosis for Dalmatians with laryngeal paralysis-polyneuropathy complex is guarded to poor. The condition is believed to be inherited.

Myotonia congenita in a Chow Chow pup.

Myotonia congenita was diagnosed in a 12-week-old female Chow Chow that had hindlimb ataxia, a stiff gait, and occasional collapsing. The diagnosis was based on the history, clinical signs, electrodiagnostics, and microscopic examination of biopsied muscle specimens.

Segmental spinal fixation of a thoracic vertebral fracture in an adult chimpanzee.

Accidental trauma associated with an epileptic convulsion in a 10-year-old, male chimpanzee (Pan troglodytes) resulted in incomplete paraplegia from fracture and subluxation of T4. Dorsal laminectomy and segmental spinal fixation were used in treatment. The segmental spinal fixation consisted of sublaminar wires attached to a contoured 316L stainless steel U-rod. The chimpanzee recovered sufficient function to allow reintroduction into the chimpanzee colony at a zoological park during the 12 months after surgery and continues to do well 24 months after surgery.

Combined Kirschner-Ehmer device and dorsal spinal plate fixation technique for caudal lumbar vertebral fractures in dogs.

A new surgical technique was developed for the fixation of caudal lumbar vertebral fractures in dogs. The technique provides optimal stabilization, can be used in combination with dorsal decompression, and does not require an intact spinous process on the fractured vertebra or attachment of the fixation devices to the fractured vertebra. The fixation consists of a Kirschner-Ehmer device and dorsal spinal plates. After fracture healing, only mild sedation of the dog is needed to allow removal of the external hardware used in the fixation. The technique, its indications, and its use in 5 cases are described.

Laryngeal paralysis in immature and mature dogs as one sign of a more diffuse polyneuropathy.

Six dogs with laryngeal paralysis had clinical, electrophysiologic, and pathologic evidence of a more generalized polyneuropathy. Three of the dogs were young Dalmatians, one was a young Bouvier des Flandres, and two were older, large-breed dogs. The results of this study suggest that laryngeal paralysis in dogs may frequently be one clinical sign of an underlying, more generalized polyneuropathy. Two forms of this generalized polyneuropathy may exist: an early form, as seen in young dogs with congenital or hereditary disease, and a delayed-onset form that is usually found in older dogs with so-called idiopathic laryngeal paralysis, some of which may have hypothyroidism.

Spinal trauma. Pathophysiology and management of traumatic spinal injuries.

Spinal trauma can originate from internal or external sources. Injuries to the spinal cord can be classified as either concussive or compressive and concussive. The pathophysiologic events surrounding spinal cord injury include the primary injury (compression, concussion) and numerous secondary injury mechanisms (vascular, biochemical, electrolyte), which are mediated by excessive oxygen free radicles, neurotransmitter and electrolyte alterations in cell membrane permeability, excitotoxic amino acids, and various other biochemical factors that collectively result in reduced SCBF, ischemia, and eventual necrosis of the gray and white matter. Management of acute spinal cord injuries includes the use of a high-dose corticosteroid regimen within the initial 8 hours after trauma. Sodium prednisolone and methylprednisolone, at recommended doses, act as oxygen radical scavengers and are anti-inflammatory. Additional considerations are the stability of the vertebral column, other conditions associated with trauma (i.e., pneumothorax), and the presence or absence of spinal cord compression, which may warrant surgical therapy. Vertebral fractures or luxations can occur in any area of the spine but most commonly occur at the junction of mobile and immobile segments. Dorsal and dorsolateral surgical approaches are applicable to the lumbosacral and thoracolumbar spine and dorsal and ventral approaches to the cervical spine. Indications for surgical intervention include spinal cord compression and vertebral instability. Instability can be determined from the type of fracture, how many of the three compartments of the vertebrae are disrupted, and on occasion, by carefully positioned stress studies of fluoroscopy. Decompression (dorsal laminectomy, hemilaminectomy, or ventral cervical slot) is employed when compression of the spinal cord exists. The hemilaminectomy (unilateral or bilateral) causes less instability than dorsal laminectomy and therefore should be used when practical. The preferred approach for atlantoaxial subluxation is ventral, and the cross pinning, vertebral fusion technique is used for stabilization. Fracture luxations of C-2 are repaired with small plates on the ventral vertebral body. The thoracic and upper lumbar spine is stabilized with dorsal fixation techniques or combined dorsal spinal plate/vertebral body plate fixation. Several methods of fixation can be used with lower lumbar or lumbosacral fractures, including the modified segmental technique and the combined dorsal spinal plate/Kirschner-Ehmer technique.

Fractures and luxations of the vertebral column.

External fixation of the vertebral column is indicated to treat fractures of the caudal lumbar spine, open fractures where vertebral osteomyelitis is present or likely to occur, and vertebral fractures not easily stabilized by internal fixation alone (e.g., compression fractures, fractures including spinous processes, and articular facets). Advantages of external fixation of caudal lumbar fractures, especially in combination with dorsal fixation devices, include the following: Fracture fixation does not preclude dorsal decompression; fixation devices need not be applied directly to the fractured vertebrae; and the combined technique provides dorsal and ventral vertebral fixation, which is more stable than dorsal fixation alone. Although external fixation is not applicable to all vertebral fracture/luxations in small animals, it provides additional points of fixation for rigid stability. Animals with vertebral fracture/luxations treated with an external fixation device have tolerated the external portion well. Development of additional applications for this method of vertebral fracture repair seems warranted.

Magnetic resonance imaging.

The advent of advanced imaging techniques in veterinary diagnostics has broadened the scope of our diagnostic capabilities. MRI is one of the newer advanced imaging techniques, and its characteristics differ considerably from CT or conventional radiography. This article presents an introduction to MRI and interpretation of MR images.

New and future advanced imaging techniques.

The advances in diagnostic imaging in the past 20 years have been nothing short of phenomenal. This article presents what is available with some of the latest in state-of-the-art techniques. Included are three-dimensional techniques, vascular MRI, high-resolution three-dimensional video animation, positron emission tomography, MR spectroscopy, and other developing methodology in diagnostic imaging.

Modulation of histamine secretion from Concanavalin A-activated rat mast cells by phosphatidyl serine, calcium, cAMP, pH and metabolic inhibitors.

Concanavalin A (con A) on its own released about half the histamine of peritoneal mast cells from three strains of rats. Phosphatidyl serine (PS) potentiated the release to about 80%. The secretory process had a calcium-independent component of 15--20% at pH 7.5, which increased to a maximum of 40% at pH 6.5 and nearly disappeared at pH 8.0. It was not affected by PS. In the presence of calcium and PS release was maximal and maintained at pH 7 to 8. Inhibitors of oxidative and glycolytic metabolism and dibutyryl cAMP (db cAMP) blocked secretion. The metabolic inhibitors were active whether or not PS was present, whereas db cAMP blocked much less effectively in the presence of PS. Dibutyryl cAMP reduced calcium uptake and this action was also impaired by PS. The active state induced by con A was relatively stable compared with that induced by antigen. Mast cells with a low response to anti-IgE also reponded poorly to con A on its own, but con A plus PS was a highly effective stimulus.

Observations on the repair of distal radial osteotomies in the dog by double hook plates.

Distal radial osteotomies in the dog repaired with a double hook plate achieved early return to full function. All dogs in this study reached and maintained the desired clinical evaluation score of grade four by the third postoperative week. The average loss of total range of carpal joint motion was 36 degrees. The double hook plate provides stable fixation and can be applied with three point fixation to a bone segment as short as 12 mm.