Functional electrical stimulation following nerve injury in a large animal model.

INTRODUCTION: Controversy exists over the effects of functional electrical stimulation (FES) on reinnervation. We hypothesized that intramuscular FES would not delay reinnervation after recurrent laryngeal nerve (RLn) axonotmesis. METHODS: RLn cryo-injury and electrode implantation in ipsilateral posterior cricoarytenoid muscle (PCA) were performed in horses. PCA was stimulated for 20 weeks in eight animals; seven served as controls. Reinnervation was monitored through muscle response to hypercapnia, electrical stimulation and exercise. Ultimately, muscle fiber type proportions and minimum fiber diameters, and RLn axon number and degree of myelination were determined. RESULTS: Laryngeal function returned to normal in both groups within 22 weeks. FES improved muscle strength and geometry, and induced increased type I:II fiber proportion (p = 0.038) in the stimulated PCA. FES showed no deleterious effects on reinnervation. DISCUSSION: Intramuscular electrical stimulation did not delay PCA reinnervation after axonotmesis. FES can represent a supportive treatment to promote laryngeal functional recovery after RLn injury. Muscle Nerve 59:717-725, 2019.

Measurement of plasma cardiac troponin I concentration by use of a point-of-care analyzer in clinically normal horses and horses with experimentally induced cardiac disease.

OBJECTIVE: To compare cardiac troponin I (cTnI) concentrations determined by use of a point-of-care analyzer with values determined by use of a bench-top immunoassay in plasma samples obtained from clinically normal horses with and without experimentally induced cardiac disease, and to establish a reference range for plasma equine cTnI concentration determined by use of the point-of-care analyzer. ANIMALS: 83 clinically normal horses, 6 of which were administered monensin to induce cardiac disease. PROCEDURES: A blood sample was collected from each of the 83 clinically normal horses to provide plasma for analysis by use of the point-of-care analyzer; some of the same samples were also analyzed by use of the immunoassay. All 83 samples were used to establish an analyzer-specific reference range for plasma cTnI concentration in clinically normal horses. In 6 horses, blood samples were also collected at various time points after administration of a single dose of monensin (1.0 to 1.5 mg/kg) via nasogastric intubation; plasma cTnI concentration in those samples was assessed by use of both methods. RESULTS: The analyzer-specific reference range for plasma cTnI concentration in clinically normal horses was 0.0 to 0.06 ng/mL. Following monensin treatment in 5 horses, increases in plasma cTnI concentration determined by use of the 2 methods were highly correlated (Pearson correlation, 0.83). Peak analyzer-determined plasma cTnI concentrations in monensin-treated horses ranged from 0.08 to 3.68 ng/mL. CONCLUSIONS AND CLINICAL RELEVANCE: In horses with and without experimentally induced cardiac disease, the point-of-care analyzer and bench-top immunoassay provided similar values of plasma cTnI concentration.

Equine laryngoplasty sutures undergo increased loading during coughing and swallowing.

OBJECTIVES: To report (1) the force required on a single laryngoplasty suture to achieve optimal abduction of the left arytenoid cartilage, (2) peak forces experienced by the suture during induced swallowing and coughing, and during 24-hour resting activity in a stall, and (3) peak forces during induced swallowing and coughing after left recurrent laryngeal nerve blockade. STUDY DESIGN: Experimental study. ANIMALS: Horses (n=8). METHODS: Each laryngoplasty suture was instrumented with an E-type buckle force transducer to measure the force required for optimal intraoperative left arytenoid cartilage abduction. This was correlated with abduction observed postoperatively. Change in suture force from baseline was measured during induced coughing and swallowing, and during normal stall activity. RESULTS: Optimal intraoperative arytenoid abduction was achieved with a mean (±SD) force of 27.6±7.5 N. During saline-induced swallowing and coughing mean force on the suture increased by 19.0±5.6 N (n=233 measurements; 7 horses) and 12.1±3.6 N (n=31; 4 horses), respectively. Sutures underwent increased loading a mean of 1152 times in 24 hours. No change in suture force was observed with respiratory rhythm. CONCLUSION: Swallowing increases laryngoplasty suture force to a greater extent than coughing.

Role of the hypoglossal nerve in equine nasopharyngeal stability.

The equine upper airway is highly adapted to provide the extremely high oxygen demand associated with strenuous aerobic exercise in this species. The tongue musculature, innervated by the hypoglossal nerve, plays an important role in airway stability in humans who also have a highly adapted upper airway to allow speech. The role of the hypoglossal nerve in stabilizing the equine upper airway has not been established. Isolated tongues from eight mature horses were dissected to determine the distal anatomy and branching of the equine hypoglossal nerve. Using this information, a peripheral nerve location technique was used to perform bilateral block of the common trunk of the hypoglossal nerve in 10 horses. Each horse was subjected to two trials with bilateral hypoglossal nerve block and two control trials (unblocked). Upper airway stability at exercise was determined using videoendoscopy and measurement of tracheal and pharyngeal pressure. Three main nerve branches were identified, medial and lateral branches and a discrete branch that innervated the geniohyoid muscle alone. Bilateral hypoglossal block induced nasopharyngeal instability in 10/19 trials, and none of the control trials (0/18) resulted in instability (P<0.001). Mean treadmill speed (+/-SD) at the onset of instability was 10.8+/-2.5 m/s. Following its onset, nasopharyngeal instability persisted until the end of the treadmill test. This instability, induced by hypoglossal nerve block, produced an expiratory obstruction similar to that seen in a naturally occurring equine disease (dorsal displacement of the soft palate, DDSP) with reduced inspiratory and expiratory pharyngeal pressure and increased expiratory tracheal pressure. These data suggest that stability of the equine upper airway at exercise may be mediated through the hypoglossal nerve. Naturally occurring DDSP in the horse shares a number of anatomic similarities with obstructive sleep apnea. Study of species with extreme respiratory adaptation, such as the horse, may provide insight into respiratory functioning in humans.

Effect of a tart cherry juice blend on exercise-induced muscle damage in horses.

OBJECTIVE: To evaluate whether administering a tart cherry juice blend (TCJB) prior to exercise would reduce skeletal and cardiac muscle damage by decreasing the inflammatory and oxidative stress response to exercise in horses. ANIMALS: 6 horses. PROCEDURES: Horses were randomly allocated into 2 groups in a crossover study with a 2-week washout period and orally administered either TCJB or a placebo solution (1.42 L, twice daily) in a double-masked protocol for 2 weeks prior to a stepwise incremental exercise protocol. Horses were tested for serum activities of creatine kinase and aspartate aminotransferase (AST) and concentrations of cardiac troponin I (cTnI), thiobarbituric acid reactive substances (TBARS; an indicator of oxidative stress), and serum amyloid A (SAA; an indicator of inflammation). To ensure that treatment would not result in positive results of an equine drug-screening protocol, serum samples obtained from each horse prior to and after 2 weeks of administration of TCJB or the placebo solution were tested. RESULTS: All horses had negative results of drug screening at both sample times. The exercise protocol resulted in a significant increase in TBARS concentration, SAA concentration, and serum AST activity in all horses. Administration of TCJB or placebo solution was not associated with an effect on malondialdehyde or SAA concentrations. However, administration of TCJB was associated with less serum activity of AST, compared with administration of placebo solution. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of TCJB may diminish muscle damage induced by exercise.

In vitro model for testing novel implants for equine laryngoplasty.

OBJECTIVE: To develop an in vitro laryngeal model to mimic airflow and pressures experienced by horses at maximal exercise with which to test laryngoplasty techniques. STUDY DESIGN: Randomized complete block. SAMPLE POPULATION: Cadaveric equine larynges (n=10). METHODS: Equine larynges were collected at necropsy and a bilateral prosthetic laryngoplasty suture was placed with #5 Fiberwire suture to achieve bilateral maximal arytenoid abduction. Each larynx was positioned in a flow chamber and subjected to static flow and dynamic flow cycling at 2 Hz. Tracheal pressure and flow, and pressure within the flow chamber were recorded at a sampling frequency of 500 Hz. Data obtained were compared with the published physiologic values for horses exercising at maximal exercise. RESULTS: Under static flow conditions, the testing system produced inspiratory tracheal pressures (mean+/-SEM) of -33.0+/-0.98 mm Hg at a flow of 54.48+/-1.8 L/s. Pressure in the flow chamber was -8.1+/-2.2 mm Hg producing a translaryngeal impedance of 0.56+/-0.15 mm Hg/L/s. Under dynamic conditions, cycling flow and pressure were reproduced at a frequency of 2 Hz, the peak inspiratory (mean+/-SEM) pharyngeal and tracheal pressures across all larynges were -8.85+/-2.5 and -35.54+/-1.6 mm Hg, respectively. Peak inspiratory flow was 51.65+/-2.3 L/s and impedance was 0.57+/-0.06 mm Hg/L/s. CONCLUSIONS: The model produced inspiratory pressures similar to those in horses at maximal exercise when airflows experienced at exercise were used. CLINICAL RELEVANCE: This model will allow testing of multiple novel techniques and may facilitate development of improved techniques for prosthetic laryngoplasty.

Measurement of digital laminar and venous temperatures as a means of comparing three methods of topically applied cold treatment for digits of horses.

OBJECTIVE: To compare effects of 3 methods of topically applied cold treatment (cryotherapy) on digital laminar and venous temperatures in horses. ANIMALS: 9 healthy adult Thoroughbreds. PROCEDURES: Thermocouples were placed in palmar digital veins and digital laminae of both forelimbs of horses. Three methods of cryotherapy were applied to the distal aspects of the limbs: wader boot (63-cm-tall vinyl boot filled with ice and water [ice slurry]), ice bag (5-L fluid bag filled with ice slurry), and a gel pack boot (boot containing frozen gel packs). Gel packs and ice slurries were replenished every hour during cryotherapy. The forelimb that received the first treatment was randomly assigned; thereafter, control and treated forelimbs were alternated for each treatment. For each treatment, temperatures were recorded every minute during 15-minute pretreatment, 2-hour treatment, and ≥ 30 minute rewarming periods. Once temperatures had returned to within 3°C below pretreatment values, the experiment was repeated in a similar manner for other cryotherapy methods. RESULTS: Digital venous temperatures were similar to laminar temperatures during each treatment. Ice bag and wader boot treatments caused similar cooling of digits. Gel boot treatment did not cause substantial cooling of digits. CONCLUSIONS AND CLINICAL RELEVANCE: Ice bag treatment caused laminar and digital venous cooling equivalent to that of wader boot treatment. Cryotherapy by use of 5-L fluid bags with an ice slurry may be a readily available, practical, and efficient method for prevention of laminitis in horses. Digital laminar and venous temperatures were similar in forelimbs of horses before and during cryotherapy.

Functional electrical stimulation of intrinsic laryngeal muscles under varying loads in exercising horses.

Bilateral vocal fold paralysis (BVCP) is a life threatening condition and appears to be a good candidate for therapy using functional electrical stimulation (FES). Developing a working FES system has been technically difficult due to the inaccessible location and small size of the sole arytenoid abductor, the posterior cricoarytenoid (PCA) muscle. A naturally-occurring disease in horses shares many functional and etiological features with BVCP. In this study, the feasibility of FES for equine vocal fold paralysis was explored by testing arytenoid abduction evoked by electrical stimulation of the PCA muscle. Rheobase and chronaxie were determined for innervated PCA muscle. We then tested the hypothesis that direct muscle stimulation can maintain airway patency during strenuous exercise in horses with induced transient conduction block of the laryngeal motor nerve. Six adult horses were instrumented with a single bipolar intra-muscular electrode in the left PCA muscle. Rheobase and chronaxie were within the normal range for innervated muscle at 0.55±0.38 v and 0.38±0.19 ms respectively. Intramuscular stimulation of the PCA muscle significantly improved arytenoid abduction at all levels of exercise intensity and there was no significant difference between the level of abduction achieved with stimulation and control values under moderate loads. The equine larynx may provide a useful model for the study of bilateral fold paralysis.