Tissue-specific expression of the human neuropeptide Y gene in transgenic mice.

Neuropeptide Y (NPY) is the most abundant neuropeptide detected in the mammalian brain, and is found throughout the central and peripheral nervous system. This peptide is a proposed regulator of appetite, blood pressure, and pituitary hormone release. Previous experiments have demonstrated the ability of 5' sequences within the human NPY gene to promote transcription in cultured neuronal cells. To identify sequences in this gene that regulate tissue-specific expression, a NPY/CAT fusion gene, containing approximately 850 bp of NPY sequences, was microinjected into fertilized mouse ova. Five lines of transgenic mice were derived from these ova and several tissues from mice of each line were tested for transgene expression using the CAT assay. One line demonstrated X-chromosome-linked transmission of the transgene while the other lines demonstrated autosomally-linked transmission. Three lines demonstrated transgene expression with significant levels of CAT activity detectable only in tissues which have been shown to express endogenous NPY. One autosomally-linked line did not demonstrate significant levels of transgene activity because the transgene appeared to have undergone structural alteration during genomic integration. No transgene activity was detected in either male of female mice from the X-linked line, suggesting a positional regulation of the transgene locus other than X-inactivation in this line. The present research demonstrated the NPY regulatory sequences included in pCATNPY delta 796 sufficiently directed tissue-appropriate gene expression in transgenic mice.

Cytogenetic effects of diethylstilbestrol-diphosphate (DES-dp) on mouse bone marrow monitored by the micronucleus test.

6 dosages of diethylstilbestrol-diphosphate (DES-dp), ranging from 0.01 to 500 mg per kg of body weight were compared to saline and phosphate buffered saline (negative controls) and two dosages of cyclophosphamide (positive control) in the micronucleus test with 115 ICR mice. DES-dp failed to generate a significant increase in micronucleated polychromatic erythrocytes over negative controls. Cyclophosphamide produced a dose-related increase in micronuclei similar to previously published reports. Iit was therefore determined that the micronucleus test did not detect the types of chromosomal changes known to be generated by DES-dp and DES.

Effects of dexamethasone and chlorpromazine treatment on X-Y dissociation and multinucleated giant cell formation in hyperthermic mice.

The effects of anti-stress drugs on X-Y dissociation and multinucleated giant cell formation in the testes of hyperthermic mice were determined for the possible use of such drugs in animal production. Mice were injected with dexamethasone 1 mg/kg or 2 mg/kg, chlorpromazine 1 mg/kg or 4 mg/kg, or 0.85% saline, and heat stressed for 4 d at 35 +/- 1 C and 65 +/- 1 % relative humidity. Mice were killed 5 d after stress, and the testes processed for observation of meiotic chromosomes and testicular histology. Heat stress caused a significant increase in X-Y bivalent dissociation in diakinesis-metaphase I spermatocytes and a significant increase in formation of multinucleated giant cells. Drug treatment increased X-Y dissociation but had no significant effect on giant cell formation.

The effect of pregnant mare serum gonadotropin on mouse oocytes as monitored at metaphase II.

Ovulated oocytes were collected from random-bred, 7-12 week old ICR mice injected with 0, 3, or 6 i.u. pregnant mare serum gonadotropin (PMSG). Analyses of 872 metaphase figures from 87 females did not show a significant increase in chromosomal imbalance with PMSG treatment. A tendency toward ovum fragmentation was noted with an increase in PMSG dose.

Cytogenic analysis of preimplantation blastocysts from prepuberal gilts treated with gonadotropins.

Metaphase chromosomes were examined in 10-day-old embryos collected from prepuberal gilts that had been induced to ovulate, using a combination of 200 IU of human chorionic gonadotropin (HCG) and 400, 600, or 800 IU of pregnant mare serum gonadotropin (PMSG) followed 72 hours later by 500 IU of HCG. A repeatable, reliable method was developed for the processing of early porcine embryos in the hatched spherical blastocyst stage for cytogenetic analysis. Of 170 embryos recovered and processed from 21 gilts, 169 yielded analyzable metaphase cells for determination of somatic cell chromosome number for an overall success rate of 99.4%. All embryos were diploid (2n = 38) in chromosomal constitution, and although isolated tetraploid (4n = 76) and octaploid (8n = 152) cells were found, true triploid or other polyploid embryos were not present. The absence of these aberrations indicated that at the dosage levels tested, the hormone PMSG did not cause an increase in occurrence of polyploidy in porcine blastocysts present at day 10 of gestation. Analyses showed no significant (P less than 0.05) increase in numbers of corpora lutea or embryos at day 10 of gestation in gilts treated with these PMSG dosage levels.

Electrodiagnostic evaluation of peripheral nerve function in rheas and barred owls.

OBJECTIVE: To establish reference values for electrodiagnostic evaluation of peripheral nerve function in birds. ANIMALS: 6 rheas and 6 barred owls. PROCEDURE: Birds were anesthetized with propofol or isoflurane in oxygen. Using a computer-based electromyograph system and needle electrodes for stimulation and recording, electromyography (EMG) was performed on the pectoral, biceps brachialis, and gastrocnemius muscles, and evoked EMG was performed on the tibial and ulnar nerves. Motor nerve conduction velocity (MNCV) was calculated. Repetitive stimulation was performed on these 2 nerves. Late F waves were recorded for each nerve, when possible. RESULTS: Activity was evident during insertion of the electrodes, but muscles tested were electrically quiescent after spontaneous EMG. Motor nerve conduction velocity was faster in the tibial nerve than ulnar nerve but did not differ significantly between species. Mean +/- SEM MNCV was 132.3+/-7.8 m/s for the tibial nerve and 59.7+/-7.8 m/s for the ulnar nerve. A significant difference was not observed in responses at the fourth or ninth stimulation during repetitive stimulation. Subsequent to the initial stimulation, amplitudes were +/-22.7% of the initial motor potential amplitude. Recorded F waves were inconsistent, which may have been associated with technique. CONCLUSIONS AND CLINICAL RELEVANCE: Reference range (mean +/-2 SEM) for MNCV was 34.1 to 75.3 m/s for the ulnar nerve and 116.7 to 147.9 m/s for the tibial nerve in barred owls and rheas. After repetitive stimulation, motor potential amplitudes may be +/-22.7% of the initial amplitude response.

Neurotoxicologic effects of the nonionic contrast agent iopamidol on the leptomeninges of the dog.

The effect of iopamidol on the leptomeninges was tested and compared with that of metrizamide and normal saline solution in 18 dogs. Pathologic and clinical effects were evaluated at 24 hours and 14 days after cisternal injection of iopamidol, metrizamide, or normal saline solution. Pathologic changes were evaluated by microscopic examination of serial CSF samples and of sections of brain and spinal cord with the leptomeninges intact. Clinical changes were subjectively evaluated. Electromyograms and EEG were performed on each dog after physical and neurologic examination. There were no changes seen in neurologic status, electromyogram, or EEG in any of the dogs immediately after subarachnoid injection nor at 24 hours or 14 days later. Pathologic changes were limited to mild, moderate, or severe patchy hemorrhagic leptomeningitis seen at 24 hours after iopamidol or metrizamide was injected. The severity of changes were judged to be similar with both these agents. The CSF analysis and histologic evaluation of brain and spinal cord sections revealed a neutrophilic response to iopamidol and a mononuclear response to metrizamide. These findings indicate that iopamidol has minimal neurotoxicologic effect on the leptomeninges and therefore has merit as a myelographic agent.

Comparison of plasma benzodiazepine concentrations following intranasal and intravenous administration of diazepam to dogs.

OBJECTIVE: To determine whether plasma concentrations of benzodiazepines (BDZ) in dogs following intranasal (IN) administration of diazepam are comparable to concentrations following IV administration. ANIMALS: 6 (4 male, 2 female) healthy adult Greyhounds. PROCEDURE: Dogs were randomly assigned to 2 groups of 3 dogs in a crossover design. Diazepam (0.5 mg/kg of body weight) was administered intravenously to dogs in group 1 and intranasally to dogs in group 2. Blood was collected from the jugular vein of each dog into tubes containing lithium heparin before and 3, 6, 9, 12, 15, 20, 30, 60, 120, 240, and 480 minutes following diazepam administration. After a 4-day washout period, dogs in group 1 received diazepam intranasally, dogs in group 2 received diazepam intravenously, and blood was again collected. Plasma concentration of BDZ was determined by use of a fluorescence polarization immunoassay. RESULTS: Mean (+/- SD) peak plasma concentration of BDZ following IV administration (1,316 +/- 216 microg/L) was greater than that following IN administration (448 +/- 41 microg/L). Time to peak concentration was < or = 3 minutes following IV administration and 4.5 +/- 1.5 minutes following IN administration. Mean bioavailability of BDZ following IN administration was 80 +/- 9%. CONCLUSIONS AND CLINICAL RELEVANCE: Diazepam is rapidly and efficiently absorbed following IN administration of the parenteral formulation. Plasma concentrations match or exceed the suggested therapeutic concentration (300 microg/L). Intranasal administration of diazepam may be useful for treatment of seizures in dogs by owners or when intravenous access is not readily available.

Electromyography in the localization of spinal cord and nerve root neoplasia in dogs and cats.

In 8 cases (6 dogs and 2 cats) of spinal cord and nerve root neoplasia, electromyography was used to localize the area of involvement. The locations of the lesions were confirmed at necropsy.

Neuroaxonal dystrophy of Rottweiler dogs.

Five Rottweiler dogs were examined because of progressive ataxia of thoracic and pelvic limbs of several months' to 4 years' duration. Hypermetria, especially of the thoracic limbs, and head incoordination and tremors were obvious features in the 3 older dogs. None of the dogs had evidence of weakness or loss of conscious proprioception. An electroencephalogram in 1 severely affected dog was normal. Electromyograms in 2 severely affected dogs were normal except for some positive sharp waves and fibrillation potentials in the interosseus muscles of all 4 feet. Cerebrospinal fluid evaluations of 2 severely affected dogs were normal. A reduced number of sensory nerve endings, compared with age-matched controls, was seen on histologic examination of a conjunctival biopsy specimen in 1 severely affected dog. An antemortem diagnosis of neuroaxonal dystrophy was suspected from the signalment, history, and neurologic examination findings. The diagnosis was confirmed on the basis of microscopic findings in 3 dogs necropsied.

Neurologic examination of sea turtles.

OBJECTIVE: To determine whether neurologic examination techniques established for use on dogs and cats could be adapted for use on sea turtles. DESIGN: Prospective controlled observational study. ANIMALS: 4 healthy Green Turtles (Chelonia mydas), 1 healthy Kemp's ridley sea turtle (Lepidochelys kempi), and 6 Green Turtles suspected to have neurologic abnormalities. PROCEDURE: Neurologic examinations were performed while sea turtles were in and out of the water and in ventral and dorsal recumbency. Mentation, general activity, head and body posture, movement and coordination, thoracic and pelvic limb movement, strength and muscle tone, and tail movement were observed. Thoracic and pelvic limb flexor reflexes and nociception, righting response, cranial nerve reflexes, clasp and cloacal reflexes, and neck, dorsal scute, cloacal and tail nociception were tested. RESULTS: Results of neurologic evaluations were consistent for healthy sea turtles. Sea turtles suspected to have neurologic abnormalities had abnormal results. CLINICAL IMPLICATIONS: Many of the neurologic examination techniques used to evaluate dogs and cats can be adapted and used to evaluate sea turtles. A standardized neurologic examination should result in an accurate assessment of neurologic function in impaired sea turtles and should help in evaluating effects of rehabilitation efforts and suitability for return to their natural environment.

Radiosurgery using a stereotactic headframe system for irradiation of brain tumors in dogs.

Radiation therapy of brain tumors in dogs typically involves administration of multiple fractions over several weeks. Fractionation is used to minimize damage to normal tissue. Radiosurgery uses multiple non-coplanar stereotactically focused beams of radiation in a series of arcs to deliver a single dose to the target with extreme accuracy. The large number of beams facilitates a high degree of conformation between the treatment area and the target tumor and allows for a steep dose gradient; the use of nonintersecting arcs minimizes exposure of normal tissue. Computed tomography with a stereotactic localizer secured to the skull allows generation of a 3-dimensional image of the target and provides accurate spatial coordinates for computerized treatment planning and delivery. Three dogs were treated with radiosurgery, using 1,000 to 1,500 cGy. A linear accelerator mounted on a rotating gantry was used to generate and deliver the radiation. Two dogs with meningiomas survived 227 and 56 weeks after radiosurgery. A dog with an oligodendroglioma survived 66 weeks. No complications were observed following the use of this technique.

Peripheral neuropathy in a turkey vulture with lead toxicosis.

Clinical, electromyographic, and pathologic findings characteristic of lead toxicosis were detected in a turkey vulture (Cathartes aura). The bird had generalized lower motor neuron dysfunction that progressed over 5 days. Electromyography revealed diffuse denervation potentials and a presumed decrement in the sciatic-tibial nerve conduction velocity. Histologic examination of peripheral nerves obtained at necropsy revealed changes that could be compatible with lead-induced neuropathy. Lead toxicosis was confirmed by determination of blood lead concentrations. Lead toxicosis causing neurologic disorders in birds has been described. However, this report emphasizes the effects of lead on the peripheral nervous system and demonstrates the use of electromyography for diagnosis of peripheral neuropathy in birds.

Cerebrospinal fluid analysis.

Accurate interpretation of cerebrospinal fluid (CSF) changes can only be made in the context of the differential diagnosis for each case. The routine analysis of CSF cell number and type as well as CSF total protein can provide information that suggests a specific mechanism or disease, but is often inconclusive. Further information obtained from CSF protein electrophoresis and immunoglobulin determination and calculation of an albumin quota and IgG index can lend additional support for the suspected mechanism of disease. Paired serum and CSF antibody titers for specific organisms can be useful to confirm the presence of a systemic or nervous system infection. Current research on detecting antibodies against nervous tissue components in CSF should result in better diagnostic capabilities and understanding of the pathophysiology of certain disorders in the future.

Influence of phenylalanine and lysine deficiencies on thyroid morphology in the chick.

A study was conducted to investigate the effects of dietary phenylalanine and lysine deficiencies on thyroid morphology in chicks fed crystalline amino acid diets. Both mean follicle diameter and epithelial cell height were significantly reduced in birds fed either of the deficient diets as compared with controls. These results agree with previous studies on serum T4 and T3 concentrations where the effects of an amino acid deficiency were general and not limited to precursors of thyroxine.

An auricular acupuncture treatment for idiopathic canine epilepsy: a preliminary report.

A new canine auricular acupuncture point for the treatment for epilepsy is introduced. The new acupoint was used with a previously reported canine auricular acupoint (van Neikerk and Eckersley, 1988) for the treatment of epilepsy in five dogs. The testimonial results of these five cases indicate that the technique is worthy of scientific investigation, and controlled research is proposed. Only testimonial evidence currently supports its application, yet this innocuous treatment is recommended for use in all cases of canine epilepsy. The traditional Chinese medical explanation for the treatment is described, and hypothetical Western physiology is presented.

Polyneuropathies of cats.

Polyneuropathies of cats have a variety of clinical presentations. Areflexic flaccid quadriparesis, or quadriplegia, progressing over a 24- to 48-hour period, may be associated with polyneuropathies, as can chronic insidiously progressive tremors and muscle weakness that wax and wane or progress slowly over weeks or months, and which can go undiagnosed for years. In addition, these neurological signs may be due to spinal cord, neuromuscular junction or muscle disorders, so the diagnosis of polyneuropathy can be a challenge even for the most astute of clinicians. Polyneuropathies may have congenital, inherited, inflammatory, metabolic and toxic causes. Sometimes the underlying aetiology is not found and a diagnosis of idiopathic polyneuropathy is made. Since the treatment and prognosis of polyneuropathies in cats vary, the purpose of this review is to assist the veterinary practitioner to recognise, appropriately manage and provide an accurate prognosis for these challenging cases.

Lipid storage myopathy in a cocker spaniel.

A six-year-old male cocker spaniel was presented to the Veterinary Medical Teaching Hospital, University of Florida, with a three-week history of generalised weakness and myalgia. Electrodiagnostic evaluation, cerebrospinal fluid analysis and thoracolumbar myelography were unremarkable. Biopsies from vastus lateralis and triceps muscles revealed numerous large lipid droplets within type 1 fibres and to a lesser degree within type 2 fibres. The resting plasma lactate was mildly increased and there was elevated urinary excretion of lactic, pyruvic and acetoacetic acids, increased urinary excretion of carnitine esters, and increased plasma alanine. This pattern of metabolite excretion is consistent with an, as yet undefined, block in oxidative metabolism.

Partial surgical removal of an intramedullary epidermoid cyst from the spinal cord of a dog.

An intramedullary space-occupying lesion in the form of an epidermoid cyst was diagnosed in a one-and-a-half-year-old flat-coated retriever. Dorsal laminectomy and durotomy were performed in order to establish the diagnosis followed by excision of one third of the cyst. The remaining cystic tissue that was intimately attached to the spinal cord parenchyma was left in place in order to avoid further damage to the nervous tissue. The dog's neurological status improved dramatically after the surgery, but deteriorated four months later due to recurrence of the cyst.

Spinal meningeal sarcoma in a rottweiler puppy.

An 11-week-old rottweiler puppy was presented for evaluation of progressive paraparesis, urinary and faecal incontinence. Radiography and myelography revealed widening of the vertebral canal and remodelling of several lumbar vertebrae associated with an intramedullary spinal mass. Magnetic resonance imaging revealed an extensive, heterogeneous mass involving the lumbosacral spinal cord. Histopathological examination indicated a meningeal sarcoma with very varied cytological architecture.