Study of the efficacy, biodistribution, and safety profile of therapeutic gutless adenovirus vectors as a prelude to a phase I clinical trial for glioblastoma.

Glioblastoma multiforme (GBM) is the most common and most aggressive primary brain tumor in humans. Systemic immunity against gene therapy vectors has been shown to hamper therapeutic efficacy; however, helper-dependent high-capacity adenovirus (HC-Ad) vectors elicit sustained transgene expression, even in the presence of systemic anti-adenoviral immunity. We engineered HC-Ads encoding the conditional cytotoxic herpes simplex type 1 thymidine kinase (TK) and the immunostimulatory cytokine fms-like tyrosine kinase ligand 3 (Flt3L). Flt3L expression is under the control of the regulatable Tet-ON system. In anticipation of a phase I clinical trial for GBM, we assessed the therapeutic efficacy, biodistribution, and clinical and neurotoxicity with escalating doses of HC-Ad-TetOn-Flt3L + HC-Ad-TK in rats. Intratumoral administration of these therapeutic HC-Ads in rats bearing large intracranial GBMs led to long-term survival in approximately 70% of the animals and development of antiglioma immunological memory without signs of neuropathology or systemic toxicity. Systemic anti-adenoviral immunity did not affect therapeutic efficacy. These data support the idea that it would be useful to develop HC-Ad vectors further as a therapeutic gene-delivery platform to implement GBM phase I clinical trials.

Nasal strips do not affect pulmonary gas exchange, anaerobic metabolism, or EIPH in exercising Thoroughbreds.

The present study was carried out to examine whether nasal strip application would improve the exercise-induced arterial hypoxemia and hypercapnia, diminish anaerobic metabolism, and modify the incidence of exercise-induced pulmonary hemorrhage (EIPH) in horses. Two sets of experiments, control and nasal strip experiments, were carried out on seven healthy, sound, exercise-trained Thoroughbred horses in random order, 7 days apart. Simultaneous measurements of core temperature, arterial and mixed venous blood gases/pH, and blood lactate and ammonia concentrations were made at rest, during submaximal and near-maximal exercise, and during recovery. In both treatments, whereas submaximal exercise caused hyperventilation, near-maximal exercise induced significant arterial hypoxemia, desaturation of Hb, hypercapnia, and acidosis. However, O2 content increased significantly with exercise in both treatments, while the mixed venous blood O2 content decreased as O2 extraction increased. In both treatments, plasma ammonia and blood lactate concentrations increased significantly with exercise. Statistically significant differences between the control and the nasal strip experiments could not be discerned, however. Also, all horses experienced EIPH in both treatments. Thus our data indicated that application of an external nasal dilator strip neither improved the exercise-induced arterial hypoxemia and hypercapnia nor diminished anaerobic metabolism or the incidence of EIPH in Thoroughbred horses performing strenuous exercise.

Pulmonary vascular pressures of thoroughbred horses exercised 1, 2, 3 and 4 h after furosemide administration.

Furosemide premedication of horses 4 h prior to exercise significantly attenuates exercise-induced pulmonary capillary hypertension which may help diminish the severity of exercise-induced pulmonary haemorrhage. As pulmonary hemodynamic effects of furosemide may be mediated via a reduction in plasma volume (which is most pronounced 15-30 min postfurosemide administration, with plasma volume recovering thereafter), we hypothesized that administration of furosemide at intervals shorter than 4 h before exertion may be more effective in attenuating the exercise-induced rise in pulmonary capillary blood pressure. Thus, our objective was to determine whether furosemide-induced attenuation of exercise-induced pulmonary arterial, capillary and venous hypertension would be enhanced when the drug is administered at intervals shorter than 4 h before exercise. Using established techniques, right atrial, and pulmonary arterial, capillary and wedge (venous) pressures were ascertained in seven healthy, sound, exercise-trained Thoroughbred horses in a randomized split-plot experimental design. Measurements were made at rest and during exercise performed at maximal heart rate (217 +/- 3 beats/min) in the control (no medications) experiments and following furosemide administration (250 mg intravenously (i.v.)) at 1, 2, 3 and 4 h before exercise. Sequence of treatments was randomized and 7 days were allowed between experiments on each horse. Although furosemide administration in the four treatment groups caused only insignificant changes in the pulmonary arterial, capillary and wedge pressures of standing horses, furosemide-induced reduction in mean right atrial pressure achieved statistical significance in the 2 h postfurosemide experiments. In the control studies, exercise was attended by statistically significant increments in mean right atrial, as well as pulmonary arterial, capillary and wedge pressures. Although exercise in each of the four furosemide experiments was also attended by significant increments in right atrial as well as pulmonary vascular pressures, in the 1, 2 and 3 h postfurosemide experiments, mean right atrial pressure increased to a significantly lower value than in the control study. Exercise-induced changes in pulmonary vascular pressures in the 1 h postfurosemide experiments were not different from the pressures in the control study. There was a significant attenuation of exercise-induced pulmonary capillary and venous hypertension in the 2, 3 and 4 h postfurosemide experiments, but significant differences among these treatments were not found. Thus, these data did not support the contention that administration of furosemide at intervals shorter than 4 h before exercise is more effective in attenuating exercise-induced pulmonary capillary or venous hypertension in Thoroughbred horses.

Short- and long-term survival and prevalence of postoperative ileus after small intestinal surgery in the horse.

The records of 74 horses that recovered from anaesthesia after surgery for a small intestinal lesion from 1994 to 1999 were reviewed. Sixty-three horses (85%) had a strangulating lesion and 43 of these (68%) had a resection and anastomosis. Four of 11 horses (36%) without a strangulating lesion had a resection and anastomosis. Sixty-three horses (85%) survived to discharge, with a survival rate of 53/63 in horses with a strangulating lesion (84%) and 10/11 (91%) in others. For all lesions, short-term survival for all end-to-end anastomoses (91%; 21/23) and for no resection (92%; 23/25) were superior (P < 0.05) to survival for jejunocaecal anastomosis (76%; 19/25). Fourteen horses (19%) had a repeat abdominal surgery during hospitalisation; 9 of these (64%) survived short-term. Postoperative ileus developed in 7/70 horses (10%) after surgery for a problem other than proximal enteritis, and all had a strangulating lesion. Postoperative ileus (POI) was more likely after a jejunocaecostomy than after other procedures, and did not develop after a jejunojejunostomy. Survival > 7 months was 52/69 (75%) and for > 12 months was 39/57 (68%). The estimated prevalence of adhesions was 13%. Short-term survival was poorest in horses that had a jejunocaecostomy, but long-term survival was less affected by the anastomosis used. The sharpest decline in survival was during the first postoperative week and postoperative mortality then declined over time after surgery. A postoperative protocol that allowed early postoperative feeding was well tolerated. The results confirm that the overall prognosis after small intestinal surgery in horses is improved over earlier findings.

Caesarean section and other methods for assisted delivery: comparison of effects on mare mortality and complications.

Data from 116 mares that had caesarean section or vaginal delivery at 2 university hospitals were analysed in 5 groups, as follows: dystocia corrected by caesarean section, Group DCS (n = 48); elective caesarean section, Group ECS (n = 10); caesarean section concurrently with colic surgery, Group CCS (n = 8); assisted vaginal delivery, Group AVD (n = 22); and controlled vaginal delivery under general anaesthesia, Group CVD (n = 28). Survival rate in all mares that had caesarean section, excluding Group CCS, was 88% (51/58). All mares in Group ECS survived and Group CCS had the lowest survival rate (38%). In 98 mares with dystocia, Groups DCS (15%) and AVD (14%) had significantly lower (P<0.05) mortality rates than Group CVD (29%). There were no differences between groups for duration of dystocia. The placenta was retained in 75 (65%) of 116 mares, and for a longer period following elective caesarean section than following assisted vaginal delivery. Multiple complications (> or = 3) were recorded in 6 mares in Group CVD but not in the other groups. Of the 102 foals delivered from 98 mares with dystocia, 11 (11%) were alive at delivery and 5 (5%) survived to discharge. Survival rate for foals was 38% in Group CCS, and 90% in Group ECS. Under conditions similar to those in this study, it is calculated that caesarean section is preferable to CVD if dystocia is protracted and great difficulty and trauma is involved, even if CVD allows delivery of the foal.

An evaluation of the haemostatic suture in hysterotomy closure in the mare.

This study was designed to evaluate the haemostatic suture as a means of preventing haemorrhage from the hysterotomy in mares after caesarean section. At 2 university hospitals 1982-1994, 48 mares had caesarean section for dystocia, 10 as an elective, and 8 mares concurrently with colic surgery. The haemostatic suture was used in 31 of 66 mares (47%) and surgery period was significantly (P<0.05) shorter when it was not applied. Anaemia (PCV<30%) was recorded in 13 (22%) of 58 mares, excluding the colic group, and the haemostatic suture did not after this proportion of mares that had anaemia. Anaemia was 5 times more probable following caesarean section than vaginal delivery, evidence that bleeding from the hysterotomy is a serious and common complication of caesarean section in mares. Severe uterine haemorrhage was recorded in 3 mares that had an haemostatic suture (10%) and in 2 mares that did not (6%). The latter two mares died of haemorrhage. The suture, therefore did not eliminate post operative anaemia and severe uterine haemorrhage. If omitted, the hysterotomy should be closed with a full thickness pattern that is sufficiently tight to compress vessels in the uterine wall.

Purification of two equine pepsinogens by use of high-performance liquid chromatography.

OBJECTIVES: To purify and characterize pepsinogens in equine gastric mucosa. SAMPLE POPULATION: Stomachs collected from 2 healthy horses at necropsy. PROCEDURE: After collection, stomachs were placed immediately in ice before storage at -48 C. After slow thawing, the mucosa was scraped off while the tissue was immersed in 0.1M potassium phosphate (pH 7.4) at 4 C, then was homogenized. The filtered extract was subjected to anion-exchange chromatography. Fractions that were found to contain pepsin or pepsinogen were further chromatographed. Individual fractions were tested for pepsinogen or pepsin content by monitoring proteolytic activity at pH 2 and 3, respectively. Fractions from all columns were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to confirm molecular weight of pepsinogens and pepsin. RESULTS: Two pepsinogens and at least 1 pepsin were purified from equine gastric mucosa. CONCLUSIONS: On the basis of molecular mass, equine gastric mucosa contains 2 pepsinogens. CLINICAL RELEVANCE: Results of this study will enable future development of an ELISA or radioimmunoassay for use in the diagnosis of equine gastric ulceration.

Failure of psyllium mucilloid to hasten evaluation of sand from the equine large intestine.

OBJECTIVE: To examine the efficacy of psyllium mucilloid in evacuating sand from the equine large intestine. ANIMALS: 12 clinically healthy pony geldings. PROCEDURE: Twelve ponies were assigned to 2 groups of six each. One group was treated with psyllium and the second was a control group. All ponies had an exploratory celiotomy and 10 g/kg body weight of sand was placed into the cecum. Ponies were fed a grain mixture alone at 1 g/kg (controls), a grain mixture plus psyllium pellets, each at 1 g/kg body weight (3 ponies), or fed a grain mixture and given psyllium powder by nasogastric tube at 1 g/kg body weight divided into two daily doses in 3 L of water (3 ponies). Radiographs were taken on days 1 (3 per group), 5 (all ponies), and 11 (3 per group) to monitor sand transit through the large intestine. Ponies were euthanatized 11 days after surgery. Sand was collected from the contents of the cecum, ventral colon, dorsal colon, and small colon. Dry weight of the recovered sand was compared between the two treatment groups as a percentage of the dry weight of sand placed in the cecum. RESULTS: No significant differences were detected in the mean percentage of sand recovered between the two treatment groups (P < .05), with 39.2% recovered in ponies treated with psyllium and 27.4% recovered in control ponies. CLINICAL RELEVANCE: Psyllium mucilloid had no apparent effect on sand evacuation from the equine large intestine. When intake of sand is prevented, the equine large intestine can reduce and possibly eliminate its sand burden.

Dental physical examination.

The objectives of the equine dental physical examination are to detect and quantify oral and dental disorders, to propose and carry out their treatment, and to implement management programs. The veterinarian should be able to offer a prognosis and to detail any future treatment or management plans that may be required. These objectives should take into account the cost of these procedures, and the veterinarian should be prepared to offer a cost-benefit analysis of the problem and the proposed cures.

Magnetic resonance imaging of the equine metacarpophalangeal joint: three-dimensional reconstruction and anatomic analysis.

Magnetic resonance imaging was used to examine the equine metacarpophalangeal joint. Thirty-two saggital images generated by partial volume imaging were transferred to a computer for three-dimensional reconstruction and analysis. All the tissues constituting the metacarpophalangeal joint were readily identified. The most significant increase finding regarded the soft tissues on the palmar aspect of the metacarpophalangeal joint and their interactions with the proximal sesamoid bones. The equine metacarpophalangeal joint has not previously been evaluated using 3-dimensional imaging software.

Correlation between anatomic features and low-field magnetic resonance imaging of the equine metacarpophalangeal joint.

OBJECTIVE: To expand our current knowledge and to establish limits of correlation between signal intensities of the magnetic resonance (MR) image and actual macroscopic and microscopic anatomic features of the imaged structures of the equine metacarpophalangeal joint (MCPJ). SAMPLE POPULATION: The right MCPJ was obtained from 4 adult horses that were euthanatized for reasons unrelated to the musculoskeletal system. PROCEDURE: The distal portion of the right forelimbs was collected from 4 equine cadavers. The bones were drilled to provide fixed reference points and examined by MR imaging. After imaging, the joints were sectioned for gross and histologic inspection. The MR images were aligned and correlated with digitized gross and histologic images to identify tissue types. RESULTS: Comparison of the images resulted in identification of different bone types, articular cartilage, and soft tissue structures of the equine MCPJ. CONCLUSION: Results provided relevant information regarding the appearance of the imaged tissues of the equine MCPJ. CLINICAL RELEVANCE: Although MR imaging does not have current clinical applications for equine practitioners, its wide acceptance as the imaging modality used for most human musculoskeletal derangements may aid in developing more realistic applications in equine medicine.

Effects of aging on the endodontic system, reserve crown, and roots of equine mandibular cheek teeth.

OBJECTIVE: To document age-related changes in the morphology of the endodontic system, reserve crown, and roots of equine mandibular cheek teeth. DESIGN: Equine mandibular cheek teeth from horses of various ages were compared, using radiography, x-ray computed tomography, and histologic examinations. SAMPLE POPULATION: 48 right hemi-mandibles from horses 2 to 9 years old. PROCEDURE: Hemi-mandibles were radiographed, imaged by computed tomographic reconstruction, and reformatted. Histologic examination was used to identify and correlate tissue types. RESULTS: Permanent mandibular cheek teeth of the horse, at the time of eruption, consisted of an exposed crown and a reserve crown with a widely dilated apex. The endodontic system consisted of 5 or 6 pulp horns that connected to an expansive pulp in the reserve crown, which was confluent with the primordial pulp bulb surrounding the tooth's apex. At the time of eruption, mandibular cheek teeth did not have a distinct pulp chamber, roots, or evidence of root formation. However, within 2 years after eruption, mesial and distal roots and a pulp chamber were present. A distinct pulp chamber, communicating with the pulp horns and both root pulp canals, was identifiable for 4 to 5 years from the time of root formation. The endodontic system of cheek teeth, 6 to 8 years after eruption, consisted of 2 unattached compartments, made up of a root canal, pulp chamber, and 2 or 3 pulp horns. CLINICAL RELEVANCE: The age-related morphologic changes in equine mandibular cheek teeth have important implications for application of endodontic therapy in horses.

Complications of celiotomy incisions in horses.

Complications of celiotomy incisions were evaluated retrospectively in 274 horses that survived at least 1 month after surgery, or died or were euthanatized within 1 month of surgery, as a direct result of these complications. Horses were divided into four groups; group A, a ventral median celiotomy for intestinal disease; group B, ventral median celiotomy for nonintestinal disease; group C, repair of an umbilical hernia; and group D, celiotomy in a region other than the midline. Specific incisional complications were peri-incisional edema, drainage, incisional abscess, suture sinus, and dehiscence. Incision-related complications occurred in 30% of the horses (group A, 40%; group B 18%; group C, 7%; and group D, 88%). Complications occurred more frequently in group D than group A (P = .009), which were higher than in groups B and C (P < .00001). Incisional hernia occurred in 28 of 256 (11%) horses that survived at least 4 months and were available for follow-up. Hernia formation was more common P < .00001) in horses that had other incisional complications (23 horses) than those without (5 horses). Serous or purulent incisional drainage, were more likely to be associated with hernia formation than was serosanguineous drainage or other incisional complications.

Evaluation of fetal infection and abortion in pregnant ponies experimentally infected with Ehrlichia risticii.

Fetal infectivity of Ehrlichia risticii was investigated in 19 ponies that were E risticii negative on the basis of results of an indirect fluorescent antibody (IFA) test. Thirteen pregnant ponies were infected by IV administration of E risticii between 90 and 180 days of gestation. Six pregnant ponies served as noninfected controls. Each infected pony had clinical signs of equine monocytic ehrlichiosis, was confirmed to be ehrlichemic, and developed an IFA titer to E risticii. Two infected ponies became recumbent, were unresponsive to supportive care, and were euthanatized. After recovery from clinical illness, the remaining ponies were observed throughout gestation for reproductive abnormalities. On abortion, each fetus was necropsied and tissue specimens from the liver, bone marrow, spleen, colon, and mesenteric lymph nodes were inoculated into canine monocyte cell cultures. Six infected ponies aborted at a mean 217 days of gestation, which was between postinoculation days 65 and 111. Five fetuses were recovered for evaluation, and E risticii was isolated from 4 of them. All 5 fetuses recovered had similar histologic finding, including enterocolitis, periportal hepatitis, and lymphoid hyperplasia with necrosis of the mesenteric lymph nodes and spleen. All 5 fetuses tested negative for IgG to E risticii, although 3 had low IgM titer to E risticii. The remaining 5 infected ponies had normal parturition. Presuckle IFA titer to E risticii was measured in 4 of the term foals, and results for 3 were positive. Two foals from infected ponies were monitored for 6 months and daily gain in body weight was comparable to that of a control foal. None of the control ponies became ill or seroconverted during the clinical illness phase, and none aborted throughout gestation Two control ponies seroconverted to E risticii 6 weeks before parturition. Results of this study indicate that E ristcii is a primary abortifacient under experimental conditions.

The macroscopic vascular anatomy of the equine ethmoidal area.

The vascular anatomy of the ethmoidal area in six normal horses and two normal ponies was studied using vascular-corrosion casts. The major arterial supply to the ethmoidal area stems from an intracranial source. The internal and external ethmoidal arteries anastomose on the rostral intracranial surface of the cribriform plate to form the arterial ethmoidal rete which arborizes and passes through the perforations of the cribriform plate to supply the ethmoid labyrinth. A minor arterial supply to the ventral portion of the ethmoid labyrinth stems from a small caudal nasal branch of the sphenopalatine artery. Multiple parallel venules drain the ethmoid labyrinth rostrally to its apex then join the venous drainage from the surrounding sinuses.

In vivo investigation of communication between the distal intertarsal and tarsometatarsal joints in horses and ponies.

The frequency of communication between the tarsometatarsal and distal intertarsal joints was investigated in 27 horses and 12 ponies. After the injection of diluted latex into the tarsometatarsal joints using the plantarolateral approach, the horses and ponies were walked for 200 meters. Latex was found in the distal intertarsal joints of 19 of 73 tarsi (26% communication) at necropsy. In 3 tarsi, latex was also found in the proximal intertarsal and tarsocrural joints.