Neuroblastoma directed therapy by a rational prodrug design of etoposide as a substrate for tyrosine hydroxylase.

Tumor directed cytotoxic therapy is one of the major challenges for the success of chemotherapy. In order to accomplish this goal in neuroblastoma, we rationally designed a prodrug of etoposide as substrate for tyrosine hydroxylase, a well established neuroblastoma associated enzyme. Here, we report synthesis and characterization of a 3,4 dihydroxy-phenyl carbamate derivative of etoposide. In order to demonstrate activation by tyrosine hydroxylase, the coding sequence of murine tyrosine hydroxylase was generated by reverse transcriptase-polymerase chain reaction from NXS2 neuroblastoma cells and cloned into the pRSET-A bacterial expression vector. The enzyme was expressed in Escherichia coli, characterized by Western blot and enzymatic activity was demonstrated by conversion of tyrosine into DOPA in the presence of cofactors using reversed phase high-performance liquid chromatography. Under these enzymatic conditions, we demonstrate conversion of 3,4 dihydroxy-phenyl carbamate prodrug into free etoposide. This effect was clearly mediated by the enzyme since bacteria transformed with the empty vector were ineffective of prodrug activation. Furthermore, tyrosine hydroxylase positive cells exposed to the etoposide prodrug were effectively killed in contrast to tyrosine hydroxylase negative controls. These findings demonstrate that etoposide can be designed as a prodrug substrate for tyrosine hydroxylase and thereby establish proof of concept for neuroblastoma directed enzyme prodrug therapy.

Internal fixation of a transverse scapular neck fracture in a filly.

A 3-week-old Standardbred filly had a non-weightbearing forelimb lameness caused by scapular neck fracture. The fracture was repaired with 2 dynamic compression plates placed 90 degrees to each other. A 10-hole 4.5-mm narrow dynamic compression plate was placed on the cranial aspect of the scapular spine, and a 10-hole 3.5-mm dynamic compression plate was placed caudal to the scapular spine. One year after surgery, the filly was not lame when exercising in the pasture, and muscle atrophy was not evident on the affected limb. Eighteen months after surgery, the filly was in race training with no apparent problems caused by fracture repair.

A preliminary investigation of the effect of selected electromagnetic field devices on healing of cannon bone osteotomies in horses.

The effect of electrical stimulation by means of selected electromagnetic field devices on healing of cannon bone osteotomies in horses was examined. The defects were created as 3 cm x 1 mm longitudinal osteotomies through the dorsal cortices of the mid-metacarpi/metatarsi of adult horses. This type of defect would asses bone healing in a situation similar to an acute, stable fracture of the cortex. Three electromagnetic devices of different design were tested in three different groups of horses. Healing was evaluated radiographically and histologically. Results showed that osteotomies treated with the electromagnetic devices healed similarly to untreated controls. Our conclusion is that the electromagnetic devices studied did not have a local effect on the repair process of an acute, stable, osseous defect.

Cruciate and collateral ligament failure in the equine stifle: seven cases (1975-1985).

Medical records of 7 adult horses with single or multiple ligament failure of the femorotibial ligaments were reviewed to evaluate signalment, history, diagnostic and treatment methods, outcome, and long-term follow-up information. Acute onset of lameness was recorded for most of the horses. Horses with multiple ligament injury had more severe clinical signs than did horses with single ligament injury. Horses with single ligament injury generally required manipulative tests to localize the lameness to the stifle. In all horses, radiography of the stifle assisted in the diagnosis. Horses either were euthanatized (n = 3) or were treated by stall rest (n = 4) after diagnosis. Of the 4 surviving horses, 2 became successful breeding animals, 1 was awaiting breeding, and 1 was used unsuccessfully as a breeding animal. Necropsy findings in 2 of the 3 horses euthanatized after diagnosis revealed early articular damage in 1 horse 1 day after injury, and degenerative joint disease in the other horse 1 year after injury.

Treatment of a ruptured medial collateral ligament of the stifle in a horse.

Medial collateral ligament ruptures in horses are rare and difficult to treat. Conservative treatment usually results in degenerative joint disease that causes permanent lameness. Surgical treatment may be an alternative for salvaging horses for breeding purpose.

Surgical reconstruction of a ruptured medial collateral ligament in a foal.

An avulsed carpal medial collateral ligament was diagnosed by clinical and radiographic examination in a 1-day-old foal that was unable to bear weight on the affected limb. The foal had a valgus deviation originating at the carpus and medial to lateral instability of the affected carpus. The ligament was reconstructed with screws and suture tape. Surgical reconstruction is a viable alternative to long-term casting or splinting for treatment of a ruptured carpal medial collateral ligament in a foal.

Congenital sinus cyst in a foal.

Radiography and endoscopy of a 17-day-old Standardbred foal that had right-sided facial swelling and dyspnea since birth revealed a soft tissue mass in the right nasal passage and right maxillary and frontal sinuses. A bone flap was used to expose the mass, and a fluid-filled structure was removed surgically. After surgery, the dyspnea was alleviated. The facial deformity resolved by the time the foal was 6 months old. The upper airway obstruction was absent clinically and endoscopically by the time the foal was 17 months old.

Radius fractures in the horse: a retrospective study of 47 cases.

The case records, radiographs and treatment results of 47 consecutive equine radius fractures presented to The Ohio State University Veterinary Hospital from 1975 to 1985 were examined retrospectively. Chip fractures and incomplete fractures were not considered. Both diaphyseal fractures and fractures involving the physes were included. Diaphyseal fractures were placed into three major categories: comminuted, oblique and transverse; and subclassified as proximal, mid-diaphyseal, or distal. Fractures involving a physis were classified using the Salter-Harris classification system. The horses ranged in age from one day to 19 years old. Nineteen of the patients were destroyed humanely on presentation, and 28 were treated. Twenty-four of the 28 fractures treated were repaired by open reduction and internal fixation. Success rate was related to the age of the horse and the type of fracture. Adult radius fractures have a poor prognosis. Most fractures in foals are either physeal fractures or transverse or oblique diaphyseal fractures and have an excellent to good prognosis when repaired adequately.

Perirectal abscesses in six horses.

Perirectal abscesses were diagnosed retrospectively in 6 horses treated for colic. The abscesses caused colic in the horses by producing an extraluminal obstruction that led to fecal impaction. The abscesses were diagnosed by rectal palpation and aspiration of exudate from the masses and were drained surgically. In horses in which the abscess involved abdominal organs, peritonitis developed.