Tissue-specific pharmacodynamics of 5-bromo-2'-deoxyuridine incorporation into DNA in VX2 tumor-bearing rabbits.

The thymidine analog 5-bromo-2'-deoxyuridine (BrdUrd) is felt to exert its cytotoxic effects primarily through incorporation into DNA. We have evaluated the incorporation of BrdUrd into the DNA of relevant normal tissues (bone marrow, gut mucosa, and liver) and tumor in rabbits with the VX2 tumor growing intrahepatically. Using constant i.v. infusions, steady state plasma drug concentrations ranging from 0.4 to 65.4 microM were maintained for 24 h and tissues were harvested and processed so that a sensitive gas chromatography/mass spectrometry (GC/MS) method could be used to analyze the thymine and 5-bromouracil content of hydrolyzed DNA. In all tissues, DNA incorporation showed saturating effects as plasma BrdUrd concentration was increased and, BrdUrd incorporation as a function of plasma concentration could be fitted to a Langmuir-like equation generating tissue-specific pharmacodynamic parameters: Imax for percentage thymine replacement at infinite plasma BrdUrd concentrations, and C50 for the arterial BrdUrd concentration generating incorporation that is Imax/2. At all plasma concentrations of BrdUrd the incorporation into DNA of bone marrow was greater than that observed in VX2 tumor. However, BrdUrd labeling index (with a BrdUrd monoclonal antibody) was greater in tumor than bone marrow. Thus, pharmacodynamic differences in incorporation do not result solely from cytokinetic differences between tissues. This model may prove useful in evaluating the pharmacodynamics of incorporation in studies using hepatic arterial infusion and biochemical modulation to improve selectivity.

Inherited necrotizing myelopathy of Afghan hounds.

Eleven examples of a naturally occurring, adolescent-onset spinal cord necrosis in Afghan Hounds were studied. Unusual and characteristic spinal cord lesions were necrosis and small vessel proliferation in the dorsal and ventro-medial spinal cord white matter extending from caudal cervical segments to cranial lumbar segments. Spinal cord gray matter and dorsal and ventral roots were preserved. Four dogs had histologically similar lesions in the superior olivary nucleus. The cause of these lesions was not determined by histopathologic or ultrastructural examination. Chi-square analysis of pooled sibships indicates a simple autosomal recessive mode of inheritance.

Laryngeal paralysis-polyneuropathy complex in young Rottweilers.

Five Rottweiler puppies from 3 unrelated litters developed inspiratory stridor at 11-13 weeks of age. Physical examination disclosed tetraparesis in all dogs, and bilateral lenticular cataracts in 4 dogs. Laryngeal examination under light anesthesia showed laryngeal paralysis in all dogs. Electrodiagnostic testing revealed denervation potentials in the distal appendicular muscles of 4 dogs tested and in the intrinsic laryngeal muscles of 2 dogs tested. Motor nerve conduction velocity was slightly low in 1 dog. Neurogenic muscular atrophy was found in distal appendicular muscles (n = 3) and intrinsic laryngeal muscles (n = 2), and degenerative changes were found in peripheral nerves (n = 3) and recurrent laryngeal nerves (n = 2). No abnormalities were detected in the spinal cord, spinal nerve roots, or ganglia of 3 dogs autopsied. The clinical, electrophysiologic, and histopathologic findings support a diagnosis of polyneuropathy and resemble the finding reported in young Dalmatians. Young dogs with laryngeal paralysis should be evaluated neurologically to rule out a more generalized polyneuropathy. The condition is suspected to be hereditary in nature and the prognosis is poor.

Familial reflex myoclonus in Labrador Retrievers.

Three 6-week-old male purebred yellow Labrador Retrievers were presented with intermittent stimulus-sensitive contractions of the appendicular and axial muscles. Five littermates were normal, although the grandsire of the affected litter had sired 2 previous litters containing similarly affected pup. Although alert and responsive, the affected dogs appeared decerebrate with extensor rigidity and opisthotonus during handling. During severe episodes, respiratory distress was observed. Generalized contractions were initiated by voluntary movements, but at rest the muscles relaxed. Neurologic deficits were not detected, although efforts to elicit segmental reflexes, assess muscle tone, or assist walking resulted in generalized stiffness. Electromyograms from the semitendinous muscles (musculi semitendinosus) had increased motor unit amplitude (up to 5,000 microV) with polyphasic action potentials. There were no myotonic discharges. A reduced interference pattern was seen. A single tactile stimuli of the distal limb resulted in 3 responses, each lasting less than or equal to 10 ms at latencies of 0 to 12 ms, 20 to 30 ms, and 35 to 40 ms, characteristic for reflex myoclonus. The motor nerve conduction velocities were normal for age (32 to 35 ms). Age-matched control dogs had motor unit action potentials of 100 to 200 microV and single compound motor unit discharges to single tactile stimuli. Therapeutic trials with diazepam and clonazepam produced minimal effects on the muscle contractions. Values of urinalyses, complete blood cell counts, and serum chemistries were within normal limits. Frozen muscle section biopsy findings, including enzyme histochemical assessment of muscle fiber types, were normal.(ABSTRACT TRUNCATED AT 250 WORDS)

Peripheral vestibular syndrome in the dog: a review of 83 cases.

Clinical data and follow-up evaluations of 83 cases of peripheral vestibular syndrome in the dog were reviewed. The absence of physical abnormalities of the tympanic membrane and absence of radiographic changes within the tympanic bulla or petrous temporal bone were the criteria used to make the diagnosis of idiopathic benign vestibular disease. When grouped according to the cause of the syndrome, most of the dogs had been diagnosed as having either otitis media/interna (49%) or idiopathic benign vestibular disease (39%). Clinical evidence suggested that most of the cases of otitis media/interna were secondary to otitis externa.

Hereditary cerebellar cortical and extrapyramidal nuclear abiotrophy in Kerry Blue Terriers.

Ten Kerry Blue terriers from a kennel in New York and 3 from a kennel in California developed clinical signs of a progressive cerebellar disorder, with its onset between 9 and 16 weeks of age. In 9 sequential necropsies, progressive cerebellar cortical degeneration, with loss of Purkinje's cells, was demonstrated. As the disease progressed, there was bilateral symmetric degeneration of the olivary nuclei followed by degeneration of the substantia nigra and caudate nucleus, bilaterally. The California and New York dogs were related, and an autosomal recessive inheritance was proposed for the disease.

The neurologic examination.

With practice, an orderly routine, and a basic understanding of neuroanatomy, the clinician should be able to tentatively localize lesions in the nervous system. Once the lesion is localized, ancillary studies are usually necessary to identify the disease process. In difficult cases when referral is impractical, an accurate description of the findings from the neurologic examination will greatly improve the value of consultation.

Brain carbohydrate metabolism during experimental Haemophilus influenzae meningitis.

Five-day-old infant rats which acquire Haemophilus influenzae b bacteremia and meningitis after intranasal inoculation have a transient depression in weight gain (2 days), but then continue to grow at the same rate as strain U--11 inoculated controls. Brain lactate, glucose, and glycogen concentrations increase during the first 5 days of disease in infected animals. The increase in brain glycogen can be accounted for by an influx of glycogen containing polymorphonuclear leukocytes. The increased concentrations of glucose and lactate were found not to be due to a change in brain weight to dry weight ratio or the volume of entrapped blood. The mean cerebrospinal fluid (CSF) glucose concentration was higher in animals with meningitis (2.7 mM) in comparison to U-11 inoculated controls (1.8 mM). This increase in brain and CSF glucose concentration appeared secondary to an increased brain uptake of hexoses as manifested by an increased [3H]mannitol uptake. Brain lactate accumulation was not explicable from the data available. There was no evidence of cerebral cortical cellular damage because in vitro oxygen uptake and lactate production were equivalent in control and meningitic animals. The ability of the infant rat brain to maintain cerebral adenosine triphosphate (ATP) content in menigitis and the failure of CSF glucose concentration to decrease might be a reflection of the importance of alternative oxidative substrate (e.g., beta-hydroxybutyrate) to the cerebral metabolism of the developing rat brain.

Neonatal endotoxin encephalopathy.

Telencephalic white matter of the neonatal kitten frequently contained diffuse astrogliosis or focal necrosis (sometimes including the thalamus and the caudate) following a single intraperitoneal injection of Escherichia coli lipopolysaccharide. No evidence for a disseminated intravascular coagulopathy was found. Telencephalic lesions in neonatal monkey and rabbit were also hemorrhagic. Enhanced karyorrhexis of glial nuclei was presented in the telencephalic white matter of the neonatal rat. In the kitten, a delay in the generation of macrophages and hypertrophic astrocytes occurs following transient neonatal endotoxemia. Marked weight loss and temperature fluctuation are prominent systemic effects. Large hemispheric cavitary lesions are not accompanied by obvious neurological deficits in the kitten.

Thiamine deficiency encephalopathy in a specific-pathogen-free cat colony.

Thiamine deficiency encephalopathy occurred in a specific-pathogen-free cat colony where the diet consisted exclusively of commercially canned diet. This product was described by the manufacturer as a complete diet for cats. The clinical diagnosis was confirmed by the demonstration of bilaterally symmetrical hemorrhage and microvascular fibrinoid necrosis in the brain and low erythrocyte transketolase levels. Treatment of clinically ill cats with vitamin B complex was followed by recovery within 24 hours.

A multiple-well method for immunohistochemical testing of many reagents on a single microscopic slide.

Multiple wells (M-wells) have been made over tissue sections on single microscopic slides to simultaneously localize binding specificity of many antibodies. More than 20 individual 4-microliter wells over tissue have been applied/slide, representing more than a 5-fold improvement in wells/slide and a 25-fold reduction in reagent volume over previous methods. More than 30 wells/slide have been applied over cellular monolayers. To produce the improvement, previous strategies of placing specimens into wells were changed to instead create wells over the specimen. We took advantage of the hydrophobic properties of paint to surround the wells and to segregate the various different primary antibodies. Segregation was complete on wells alternating with and without primary monoclonal antibody. The procedure accommodates both frozen and paraffin sections, yielding slides which last more than a year. After monoclonal antibody detection, standard histologic stains can be applied as counterstains. M-wells are suitable for localizing binding of multiple reagents or sample unknowns (polyclonal or monoclonal antibodies, hybridoma supernatants, body fluids, lectins) to either tissues or cells. Their small sample volume and large number of sample wells/slide could be particularly useful for early screening of hybridoma supernatants and for titration curves in immunohistochemistry (McKeever PE, Shakui P, Letica LH, Averill DR: J Histochem Cytochem 36:931, 1988).

The identification of Eperythrozoon ovis in anemic sheep.

Eperythrozoon ovis, a rickettsial parasite of erythrocytes, was found in anemic lambs maintained for reproductive endocrinology research. The parasite was identified in the blood films of 13 animals in the flock of 30. The sexes were infected equally (7/16 males versus 6/14 females). The relationship between the severity of the anemia and the presence of organisms in blood was statistically significant. One animal died with severe anemia. Light, scanning electron, and transmission electron microscopy of peripheral erythrocytes revealed an extracellular organism identified as E. ovis. These findings indicate that this parasite can cause disease in sheep and therefore may interfere with biomedical research.