The effects of Aurora Kinase inhibition on thyroid cancer growth and sensitivity to MAPK-directed therapies.

Thyroid cancer is one of the deadliest endocrine cancers, and its incidence has been increasing. While mutations in BRAF are common in thyroid cancer, advanced PTC patients currently lack therapeutic options targeting the MAPK pathway, and despite the approved combination of BRAF and MEK1/2 inhibition for BRAF-mutant ATC, resistance often occurs. Here, we assess growth and signaling responses to combined BRAF and MEK1/2 inhibition in a panel of BRAF-mutant thyroid cancer cell lines. We first showed that combined BRAF and MEK1/2 inhibition synergistically inhibits cell growth in four out of six of the -BRAF-mutant thyroid cancer cell lines tested. Western blotting showed that the MAPK pathway was robustly inhibited in all cell lines. Therefore, to identify potential mechanisms of resistance, we performed RNA-sequencing in cells sensitive or resistant to MEK1/2 inhibition. In response to MEK1/2 inhibition, we identified a downregulation of Aurora Kinase B (AURKB) in sensitive but not resistant cells. We further demonstrated that combined MEK1/2 and AURKB inhibition slowed cell growth, which was phenocopied by inhibiting AURKB and ERK1/2. Finally, we show that combined AURKB and ERK1/2 inhibition induces apoptosis in BRAF-mutant thyroid cancer cell lines, together suggesting a potential combination therapy for BRAF-mutant thyroid cancer patients.

Use of a novel helical fan beam imaging system for computed tomography of the distal limb in sedated standing horses: 167 cases (2019-2020).

OBJECTIVE: To evaluate the diagnostic capabilities of a novel helical fan beam CT system used for imaging of horses with a range of clinical distal limb problems. ANIMALS: 167 horses. PROCEDURES: Medical records were reviewed of horses presented for CT of the distal limb at 2 university-based veterinary hospitals. The following data were recorded: age, sex, breed, presenting complaint, sedation used for imaging, scanning time, procedure time, other diagnostic imaging methods performed, imaging diagnosis, clinical diagnosis, and complications during imaging. RESULTS: Most horses were Thoroughbreds and Quarter Horses. Procedure times ranged from 15 to 40 minutes, with scanning completed in 15 to 45 seconds for each region of interest. The foot or pastern region was commonly scanned (88/167 [53%] horses), with navicular bone disease diagnosed in 42 of 88 (48%) horses. The fetlock region was also commonly scanned (42/167 [40%] horses), with palmar or plantar osteochondral disease diagnosed in 17 of 42 (40%) horses. Horses were compliant during scanning, and no complications with sedation or damage to the scanner occurred. A specific imaging diagnosis for the lameness was achieved more frequently with CT imaging (166/167 [99%]) than with planar digital radiography (26/58 [45%]). CLINICAL RELEVANCE: The helical fan beam CT system could be used safely to scan sedated standing horses from the carpal or tarsal region distally. Subjectively, the machine was easy to operate, allowing CT to be incorporated into lameness investigations. CT imaging was very likely to result in a clinical diagnosis in horses with distal limb lameness.

Use of a novel helical fan beam imaging system for computed tomography of the head and neck in sedated standing horses: 120 cases (2019-2020).

OBJECTIVE: To evaluate the diagnostic capabilities of a novel helical fan beam CT system used for imaging of horses with clinical problems of the head and neck. ANIMALS: 120 horses. PROCEDURES: Medical records were reviewed of horses presented for CT of the head or neck at 2 university hospitals. The following data were recorded: age, sex, breed, presenting complaint, sedation used for imaging, scanning time, procedure time, other diagnostic imaging methods performed, imaging diagnosis, clinical diagnosis, and complications during imaging. RESULTS: Quarter Horses and Warmbloods were the most common breeds, and the most common complaint was nasal discharge. The head (101/120 [84%] horses) was scanned most frequently, and the most common diagnoses were primary dental disease and a space-occupying lesion of the paranasal sinuses. Nuchal bursitis was the most common imaging diagnosis in the neck region. Procedure time ranged from 20 to 45 minutes with a scanning time of 30 to 40 seconds. No complications with horse sedation occurred, and horses tolerated scanning well. An imaging diagnosis was more frequently achieved with CT (109/120 [91%] horses) than with planar digital radiography (23/61 [38%] horses). CLINICAL RELEVANCE: The helical fan beam CT system had fast scanning times and could be used safely for routine imaging of the teeth and sinuses in horses. The caudal extent of scanning in the neck region was limited by the shape of the horse's neck and thorax.

Retrobulbar lidocaine injection via the supraorbital fossa is safe in adult horses but produces regionally variable periocular anaesthesia.

BACKGROUND: Injection techniques for retrobulbar anaesthesia are published in horses, but neither safety nor anaesthetic efficacy and duration have been evaluated objectively in vivo. OBJECTIVE: To characterise the safety and efficacy of one published technique for retrobulbar anaesthesia. STUDY DESIGN: Randomised, controlled descriptive experiment. METHODS: Unilateral retrobulbar injection with 10 mL lidocaine (2%) was performed in eight sedated adult mares. Contralateral eyes served as untreated controls. Neurophthalmic parameters, intraocular pressure (IOP), and corneal and periocular sensation were measured awake, post-sedation and at periodic time points for 24 hours following injection. Adverse effects were documented. RESULTS: Injection of 10 mL lidocaine significantly increased IOP for up to 2 hours (P < .05) maximally at 30 min (mean [95% CI]: 6.0 [2.7, 9.2] mm Hg, P < .001). Six of the eight treated eyes developed mild to moderate reversible chemosis for 2 to 24 hours. One eye developed severe chemosis and superficial corneal ulceration at 24 and 48 hours following injection respectively. Corneal sensitivity significantly decreased for 6 hours (P < .05), maximally at 10 min (-44.4 [-34.6, -54.1] mm, P < .001). Periocular sensitivity (measured as increase in applied force) significantly decreased dorsally and medially for up to 2 hours (maximal at 2 hours (367.1 [238.5, 495.7] g, P < .001, and at 30 min: 345.8 [202.6, 488.9] g, P < .001) respectively). Ventral and lateral sensitivity were not effectively decreased beyond 30 min. Optic nerve function was not consistently reduced following injection. MAIN LIMITATIONS: Investigators were not masked to the treated eye. CONCLUSIONS: Retrobulbar injection using 10 mL lidocaine is safe in normal eyes of adult horses, but carries risk in structurally compromised or glaucomatous eyes due to transient IOP increase. Reversible chemosis commonly develops 2-4h following injection, and may be severe in some horses with risk for corneal ulceration. Corneal anaesthesia is rapid and prolonged, but all periocular regions are not consistently anaesthetised. Retrobulbar injection should be combined with other local anaesthetic injections for eyelid surgeries or enucleations.

Computed tomographic assessment of brain tissue disruption and skull damage in equine cadaveric heads caused by various firearm-ammunition combinations applied as potential gunshot methods for euthanasia of horses.

OBJECTIVE: To evaluate with CT the characteristics of brain tissue disruption and skull damage in cadaveric heads of adult horses caused by each of 6 firearm-ammunition combinations applied at a novel anatomic aiming point. SAMPLE: 53 equine cadaveric heads. PROCEDURES: Heads placed to simulate that of a standing horse were shot with 1 of 6 firearm-ammunition combinations applied at an aiming point along the external sagittal crest of the head where the 2 temporalis muscles form an inverted V. Firearm-ammunition combinations investigated included a .22-caliber long rifle pistol firing a 40-grain, plated lead, solid-core or hollow-point bullet (HPB); a semiautomatic 9-mm pistol firing a 115-grain, jacketed HPB; a semiautomatic .223-caliber carbine firing a 55-grain, jacketed HPB; a semiautomatic .45-caliber automatic Colt pistol firing a 230-grain, jacketed HPB; and a 12-gauge shotgun firing a 1-oz rifled slug. Additional heads placed in a simulated laterally recumbent position were shot with the semiautomatic 9-mm pistol-HPB combination. All heads underwent CT before and after being shot, and images were evaluated for projectile fragmentation, skull fracture, and cerebrum, cerebellum, and brainstem disruption. RESULTS: Computed tomography revealed that all firearm-ammunition combinations caused disruption of the cerebrum, cerebellum, and brainstem that appeared sufficient to result in instantaneous death of a live horse. Hollow-point ammunition was as effective as solid-core ammunition with regard to brain tissue disruption. Brain tissue disruption was not affected by head positioning. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the examined firearm-ammunition combinations, when applied at a novel aiming point, appear to be reasonable options for euthanasia of horses.