Chordomas at High Prevalence in the Captive Population of the Endangered Perdido Key Beach Mouse (Peromyscus polionotus trissyllepsis).

The Perdido Key beach mouse (Peromyscus polionotus trissyllepsis) is a critically endangered subspecies of the oldfield mouse. The captive population, currently maintained by 3 Florida zoos, is entirely derived from just 3 wild-caught ancestor mice. Necropsy and histopathology revealed chordoma of the vertebral column in 38 of 88 (43%) mice. The tumors were locally expansile and invasive masses of large physaliferous (vacuolated) cells with small, round, hyperchromatic nuclei, similar to the "classic" form of chordomas described in humans. Primary tumors rarely contained small amounts of bone and cartilaginous matrix, characteristic of the "chondroid" form. Neoplastic cells contained abundant granules positive by the periodic acid-Schiff reaction. Brachyury and cytokeratin AE1/AE3 were detected in neoplastic cells by immunohistochemistry, but uncoupling protein 1 was not identified. Primary tumors occurred along the entire vertebral column--cervical, 5 of 38 (13%); thoracic, 16 (42%); lumbar, 13 (34%); and sacral, 10 (26%)--and 10 (26%) mice had multiple primary masses. Metastases to the lungs were noted in 13 of the 38 (34%) mice. Mice diagnosed with chordomas postmortem ranged from 424 to 2170 days old, with a mean of 1399 days. The prevalence of chordoma was not significantly different between males (n = 23 of 50; 46%) and females (n = 15 of 38; 39%).

Pegylated feline granulocyte colony-stimulating factor increases neutrophil levels in cats.

Neutropenia can often be corrected by treatment with granulocyte-colony stimulating factor (G-CSF) and off-label use of commercial human G-CSF (HuG-CSF) is a commonly used treatment for neutropenic animals. However, long-term HuG-CSF treatment can be associated with adverse effects, including neutropenia. Here, feline (Fe) G-CSF was produced in Pichia pastoris, pegylated (Peg) FeG-CSF and tested in cats. A randomized controlled clinical trial was conducted to evaluate the efficacy of PegFeG-CSF compared to FeG-CSF or HuG-CSF in FIV-infected (n=14), FIV-uninfected healthy cats (n=19), and in HuG-CSF-induced neutropenic cats (n=4). Daily FeG-CSF doses induced higher neutrophil production than HuG-CSF after the second week of treatment (P ⩽ 0.002). Weekly doses of PegFeG-CSF induced higher neutrophil counts and showed greater sustained activity than weekly doses of FeG-CSF. PegFeG-CSF provided the most therapeutic and sustainable neutrophil production (P<0.001) in both FIV-uninfected and FIV-infected cats, without the development of neutralizing antibodies. Conversely, all HuG-CSF-treated cats developed neutralizing antibodies, suggesting cross-reactive antibodies to endogenous G-CSF in a majority of the cases with severe neutropenia. Strikingly, when PegFeG-CSF was used to rescue cats with HuG-CSF-induced neutropenia, clinically normal neutrophil numbers returned. Thus, PegFeG-CSF appears to be a superior treatment for neutropenia in feline patients.

Cranial mediastinal liposarcoma in a horse.

A 23-year-old Anglo-Arabian mare was presented with tachypnea, dyspnea, and pitting edema of the ventral thoracic subcutis. On necropsy, a tan to red, friable, irregularly shaped mass (23 × 20 × 18 cm) occupied the cranial mediastinum. Histologically, the mass was classified as a liposarcoma and was composed of short interlacing bundles of spindle-shaped to irregularly rounded cells with discrete, variably sized, clear cytoplasmic vacuoles, which were stained with oil red O in frozen sections of formalin-fixed tissue.

Evolutionarily conserved T-cell epitopes on FIV for designing an HIV/AIDS vaccine.

This review will discuss the current state of the human HIV-1 vaccine trials including the safety consideration of vaccine composition and difficulties in determining and defining protective immunity and epitopes to HIV-1. Vaccines in animal models of lentivirus infection are compared. In particular, the findings from the prototype FIV vaccine and the HIV-1 protein immunizations studies in cats are discussed, as well as the resulting research regarding a potential HIV-1 vaccine design based on evolutionarily conserved T-cell epitopes.

Mucosal injury and inflammatory cells in response to brief ischaemia and reperfusion in the equine large colon.

REASON FOR PERFORMING STUDY: Intestinal ischaemia and reperfusion (I/R) can activate inflammatory cells in the equine colon, although effects on different types of inflammatory cells have received little attention. OBJECTIVES: To assess early mucosal injury, the reaction of mucosal neutrophils, eosinophils, mast cells and macrophages, and cyclooxygenase (COX)-1 and -2 expression in response to I/R in the equine large colon. METHODS: Large colon ischaemia was induced for 1 h (1hI) followed by 4 h of reperfusion in 6 horses, and mucosal biopsies were sampled before and after ischaemia, and after 1, 2 and 4 h of reperfusion. Semithin sections (500 nm) of epon-embedded biopsies were stained with toluidine blue for histomorphometric evaluation. The number and distribution of mucosal macrophages (CD163), neutrophils (calprotectin), eosinophils (LUNA) and mast cells (toluidine blue) were determined, and mucosal COX-1 and -2 expression was identified. RESULTS: Ischaemia caused epithelial cell and nuclear swelling (mean ± s.e. nuclear width; control: 2.7 ± 0.2 µm vs. 1hI: 4.2 ± 0.2 µm; P<0.01), subepithelial oedema (control: 0.2 ± 0.1 µm vs. 1hI: 3.2 ± 0.2 µm; P<0.01) and increased epithelial apoptosis (control: 14.3 ± 4.1 apoptotic cells/mm mucosa vs. 1hI: 60.4 ± 14.0 apoptotic cells/mm mucosa; P<0.01). COX-2 expression (P<0.01) was evident after ischaemia. Reperfusion caused paracellular fluid accumulation (control: 0.9 ± 0.1 µm vs. 1hI: 0.6 ± 0.6 µm vs. 1hI + 4hR: 1.6 ± 0.2 µm; P<0.05). Epithelial repair started at 1 h of reperfusion (P<0.001), followed by migration of neutrophils into the mucosa after 2 h (control: 72.3 ± 18.4 cells/mm(2) mucosa vs. 1hI + 2hR: 1149.9 ± 220.6 cells/mm(2) mucosa; P<0.01). Mucosal eosinophils, mast cells and macrophages did not increase in numbers but were activated. CONCLUSIONS: Epithelial injury and COX-2 expression caused by short-term hypoxia were followed by intense inflammation associated with epithelial repair during reperfusion. POTENTIAL RELEVANCE: Equine colonic mucosa subjected to a brief period of ischaemia can repair during reperfusion, despite increased mucosal inflammation.

Investigation of endothelial cells as an in vivo nidus of Anaplasma marginale infection in cattle.

Continuous culture of Anaplasma marginale in endothelial cells and the potential implications for vaccine development heightened interest in determining the importance of endothelial cells in the A. marginale life cycle. A. marginale-infection trials were performed to determine if endothelial cells are an in vivo host cell in cattle and if A. marginale from in vitro endothelial cells were infective to cattle. Adult, immunocompetent steers were infected by tick-feeding transmission and were euthanized at different points in the parasitemic cycle. Based on quantitative PCR, the tissue distribution of A. marginale DNA during peak and trough parasitemia was variable with higher quantities observed in spleen, lung, hemal nodes, and abomasum. A. marginale was not conclusively identified in tissue endothelial cells from the steers' tick-bitten dermis or post-mortem tissues using three microscopy techniques (dual indirect immunofluorescence, transmission electron microscopy, and in situ DNA target-primed rolling-circle amplification of a padlock probe). Intravenous inoculation of spleen-intact or splenectomized calves with endothelial cell culture-derived VA isolate A. marginale did not cause seroconversion or clinical anaplasmosis regardless of whether the endothelial culture-derived bacteria were inoculated as host cell-free organisms or within endothelial cells and regardless of the type of endothelial cell culture used - RF/6A primate endothelial cells or primary bovine testicular vein endothelial cells. Data presented here suggest that endothelial cells are likely not a pivotal component of the A. marginale life cycle in vivo.

Retention of cast crown copings cemented to implant abutments.

BACKGROUND: The cementation of crowns to dental implant abutments is an accepted form of crown retention that requires consideration of the properties of available cements within the applied clinical context. Dental luting agents are exposed to a number of stressors that may reduce crown retention in vivo, not the least of which is occlusal loading. This study investigated the influence of compressive cyclic loading on the physical retention of cast crown copings cemented to implant abutments. METHODS: Cast crown copings were cemented to Straumann synOcta titanium implant abutments with three different readily used and available cements. Specimens were placed in a humidifier, thermocycled and subjected to one of four quantities of compressive cyclic loading. The uniaxial tensile force required to remove the cast crown copings was then recorded. RESULTS: The mean retention values for crown copings cemented with Panavia-F cement were statistically significantly greater than both KetacCem and TempBond non-eugenol cements at each compressive cyclic loading quantity. KetacCem and TempBond non-eugenol cements produced relatively low mean retention values that were not statistically significantly different at each quantity of compressive cyclic loading. Compressive cyclic loading had a statistically significant effect on Panavia-F specimens alone, but increased loading quantities produced no further statistically significant difference in mean retention. CONCLUSIONS: Within the limitations of the current in vitro conditions employed in this study, the retention of cast crown copings cemented to Straumann synOcta implant abutments with a resin, glass ionomer and temporary cement was significantly affected by cement type but not compressive cyclic loading. Resin cement is the cement of choice for the definitive non-retrievable cementation of cast crown copings to Straumann synOcta implant abutments out of the three cements tested.

Canine coronavirus-associated puppy mortality without evidence of concurrent canine parvovirus infection.

This report presents 2 cases in which puppy fatalities were associated with canine coronavirus (CCV), but no evidence of concurrent canine parvovirus (CPV-2) disease was observed. Case 1 involved a 7-week-old, male short-haired Chihuahua, which had become lethargic 24 hours after purchase from a pet store. Within 72 hours, the puppy began to vomit, had diarrhea, and was admitted to the veterinary clinic, where it was placed on IV fluids. The parvovirus Cite test was negative. The puppy died within 12 hours of admission and was submitted for diagnostic workup. Gross pathology revealed an enteritis suggestive of CPV-2. Histopathology on intestines showed scattered dilated crypts with necrotic cellular debris and neutrophils. There was moderate depletion and necrosis of lymphoid follicles. Electron microscopy (EM) on intestinal contents was positive for coronavirus and negative for parvovirus. Immunohistochemistry (IHC) on gut sections was positive for CCV and negative for CPV-2. Case 2 was an 8-week-old, male Shih Tzu, which was admitted to the veterinary clinic exhibiting symptoms of severe gastroenteritis with abdominal pain. The referring veterinarian euthanized the puppy, and the entire body was submitted for diagnostic evaluation. Necropsy revealed a severe ileo-cecal intussusception and segmental necrotic enteritis of the small intestine. Electron microscopy of the intestinal contents was positive for coronavirus and negative for parvovirus. Immunohistochemistry on sections of affected gut were positive for CCV and negative for CPV-2. These cases emphasize the importance of pursuing a diagnosis of CCV in young puppies when CPV-2 disease has been ruled out by IHC.

Persistence of Anaplasma ovis infection and conservation of the msp-2 and msp-3 multigene families within the genus Anaplasma.

Goats which have recovered from acute Anaplasma ovis infection remain seropositive, although infected erythrocytes cannot be detected by microscopic examination. Persistence of A. ovis 17 to 21 months following experimental infection was demonstrated by PCR detection of the msp-5 gene. Quantitative analysis of persistent rickettsemia over time showed that all levels were below the limit of microscopic detection and ranged from a low of 10(2) organisms/ml to peaks of 10(6) organisms/ml. Two patterns of persistent rickettsemia were observed: the first was characterized by cyclic fluctuations at 6- to 9-week intervals, similar to the pattern described for A. marginale-infected cattle, while in the second pattern, repetitive cycles did not occur and the rickettsemia levels were relatively constant. The msp-2 and msp-3 multigene families, which provide the genetic capacity for outer membrane protein antigenic variation during persistent A. marginale rickettsemia, were identified in the A. ovis genome by Southern blot analysis, and expression of an MSP-2 homologue was confirmed by using immunoblots.

Computer generated mandibular model: surgical role.

A life-size nylon model of a traumatized mandible was produced from CT scan data by the process of laser sintering. The model was used for pre-operative planning and for production of surgical aids in order to facilitate the restoration of a large bony defect. Vascularized iliac crest bone was harvested, titanium implants placed and the bone then grafted to the mandible.

Titanium cranioplasty using 3-D computer modelling of skull defects.

The ideal prosthesis for cranial defect repair is the original bone flap, but for various reasons this is not always available. Creation of prostheses has been an inexact and difficult science, particularly for complex and cosmetically important defects. Poorly shaped prostheses are difficult to insert and secure. In addition, the resultant deformity may lead to scalp necrosis and infection. The need for precision-fitting prostheses has prompted the development of several computer-aided methods of cranioplasty plate manufacture. Defects have been modelled from CT scan data using stacked slices, computer-driven milling and selective laser sintering. We report our experience with titanium cranioplasty plates manufactured using these methods. The plates fit even the most complex defects, are easy to insert, and produce excellent cosmetic results. There has been no postoperative scalp necrosis or infection. The cost and time involved may currently preclude the routine use of computer modelling of defects, but as equipment costs and production time diminish, this may become the standard allograft cranioplasty technique.

Voltage-sensitive calcium channel development in epileptic DBA/2J mice suggests altered presynaptic function.

Aberrant synapse formation has been implicated in development and propagation of epileptic potential. Litzinger et al. (1993a) showed that omega-GVIA conotoxin may be used as a marker for synapse formation in nonepileptic mice. We conducted omega-GVIA binding in synaptosomal preparations from epileptic DBA/2J mice at different developmental ages. Binding in DBA/2J mice was compared with omega-GVIA binding in synaptosomal preparations from nonepileptic C57/B1, Swiss Webster, and AJ mice. Striking differences between these strains of mice are evident in the developmental sequence and pattern of N-type voltage-sensitive calcium channels (VSCC). In contrast to nonepileptic mice, the DBA/2J mice show a slow increase in omega-GVIA binding between postnatal days 2 and 8. This increase corresponds to onset of susceptibility to seizure in this strain. In addition to the difference in developmental sequence, DBA/2J mice have fewer binding sites for omega-GVIA throughout development, suggesting changes in channel structure or number. These data show that in DBA/2J mice development of the VSCC in brain is different from that in nonepileptic mice. This difference in development in presynaptic membranes responsible for neurotransmitter release may represent a change in synaptic activity that plays a role in epileptogenesis.

Different omega-conotoxins mark the development of Swiss Webster mouse cortex suggesting N-type voltage sensitive calcium channel subtypes.

omega-GVIA conotoxin has been used to mark presynaptic N-type voltage sensitive calcium channels (VSCC). Litzinger et al. used omega-conotoxin binding to describe a critical period of neurodevelopment in Swiss Webster mice between postnatal days (PND) 11 and 14, which appears to be important to the initiation of proper final development of the central nervous system. In this study, we compare how three different omega-conotoxins (i.e. GVIA from Conus geographus, MVIIA from Conus magus, and RVIA from Conus radiatus) mark N-type VSCC during this critical period in Swiss Webster mouse cortex. 125I-GVIA was bound to Swiss Webster mouse cortex synaptosomal membrane fractions at postnatal days 8 and 14. 125I-GVIA binding displacement curves were obtained by incubating membranes with increasing concentrations of unlabeled GVIA, MVIIA, and RVIA. Displacement curves and IC50 were calculated for each of these three omega-conotoxins, and then compared. At PND 14, GVIA, MVIIA and RVIA were able to displace greater than 95% of 125I-GVIA binding. At PND 8, however, MVIIA was only able to displace 83% of 125I-GVIA binding, and RVIA was only able to block 84%. The IC50 does not appear to change significantly during this period of development for any of the omega-conotoxins. The inability of MVIIA and RVIA to completely block 125I-GVIA binding in pre-critical period Swiss Webster cortex denotes an alteration in the composition of N-type VSCC binding sites. With this data, we have suggested the presence of subtypes of the N-type VSCC in the cortex of pre-critical period Swiss Webster mouse.

Regional differences in the critical period neurodevelopment in the mouse: implications for neonatal seizures.

The voltage-sensitive calcium channel probe 125I-omega-GVIA conotoxin has been shown to be a developmental marker in whole brain preparations of Swiss Webster mice. The present study looks more carefully at regional dissections of the mouse brain (cerebrum, cerebellum, and brain stem) at postnatal day 8 and postnatal day 16. 125I-omega-GVIA conotoxin binding, thought to be presynaptic, showed a dramatic increase between postnatal days 8 and 16 in the cerebral cortex, a decrease in the cerebellum, and no change in the brain stem. The dramatic cerebral cortex increases indicated by these binding data correspond to a critical period between postnatal day 11 and postnatal day 14 in Swiss Webster mice; during this critical period, dendrites exhibit rapid outgrowth, sensory modalities come on line, electroencephalographic patterns mature, and the cortex reaches adult proportions. This period parallels a similar initiation of electrical maturation in the 28- to 32-week neonatal human brain. We conclude from these data that the unusual clinical presentation of neonatal seizures is not just the result of immature myelin formation. It includes incomplete synapse formation linking the cortex to the brain stem.

Voltage sensitive calcium channels mark a critical period in mouse neurodevelopment.

Voltage sensitive calcium channel (VSCC) probes 125I-omega-GVIA Conotoxin (omega-GVIA), (+)-[5-methyl-3H]-PN200-110 (3H-PN200), and 3H-Nimodipine were bound to developing Swiss Webster mouse whole brain from postnatal days 3 to 24. 125I-omega-GVIA binding, thought to be presynaptic, showed a 50% increase between days 11 and 14. 3H-dihydropyridine binding, thought to be postsynaptic, showed spike patterns when measured developmentally. 3H-PN200 binding showed a > 150% increase between days 11 and 15. 3H-Nimodipine binding showed a > 100% increase between days 11 and 14. Depolarization-induced 45Ca fluxes also increased between days 8 and 16 by > 500%. The dramatic increases indicated by these binding data correspond to a critical period described by Himwich (Int. Rev. Neurobiol. 4, 117, 1962) between postnatal days 11 and 14 in Swiss Webster mice; during this critical period, dendrites exhibit rapid outgrowth, sensory modalities come on line, EEG patterns mature, and the cortex reaches adult proportions. We conclude from these data that the increase in VSCC activity parallels a critical period in the development of the central nervous system in Swiss Webster mice.

Influence of the maxillary canine on mandibular fracture.

The length of the root of the mandibular canine tooth has been considered by many authors as being a source of weakness in the mandible. It has also been suggested that a direct blow or a bending force around this tooth can result in traumatic injury. We advance a theory that implicates the maxillary canine tooth as directly contributing to the mandibular canine region fracture pattern.

The missing maxilla: restoring aesthetic balance with mandibular surgery.

Traumatic loss of large portions of the maxilla remains uncommon. Reconstruction demands careful attention to both the anatomy of the primary deformity as well as the associated secondary changes if there is to be acceptable restoration of facial aesthetics and function. In concert with rebuilding the missing maxilla, reconstruction may frequently involve repositioning the apparently undisturbed mandible.

Monocortical non-compression miniplate osteosynthesis of mandibular angle fractures.

The technique of monocortical non-compression miniplate fixation of mandibular angle fractures is reviewed. A study of our first 50 patients treated using this technique reveals that consistent reduction and stabilization of these mandibular fractures can be achieved without the requirement for intermaxillary fixation. Such results were produced with minimal postoperative morbidity.

Influence of alloy composition on the hardening of silver-tin dental amalgam.

The objective of the investigation was to examine the reactions of mercury with silver-tin alloys with compositions spanning the phase fields beta, (beta + gamma), gamma, and (gamma + Sn). The experimental methods employed include the application of light microscopy, scanning electron microscopy, and electron probe microanalysis. These techniques were used to investigate the mechanisms of reaction and to identify the nature and morphology of the reaction products formed on bulk specimens of the alloys. The progress and characteristics of the reactions that occur during hardening of amalgams prepared from powders of these alloys were monitored using a high-sensitivity dilatometer. These results were correlated with direct observations on the development of the microstructures. The reaction of mercury with the beta-phase alloy occurred rapidly and resulted in a very marked and rapid expansion during the initial stages of hardening. gamma-Phase alloys, on the other hand, reacted more slowly and contracted markedly during hardening. The behavior of amalgams made from alloys with compositions lying between these two extremes appeared to be explicable in terms of the characteristics of the separate phases from which they were constituted.