The effects of simulated gastroesophageal reflux on infant pig oropharyngeal feeding physiology.

The neural connectivity among the oral cavity, pharynx, and esophagus is a critical component of infant feeding physiology. Central integration of oral and pharyngeal afferents alters motor outputs to structures that power swallowing, but the potential effects of esophageal afferents on preesophageal feeding physiology are unclear. These effects may explain the prevalence of oropharyngeal dysphagia in infants suffering from gastroesophageal reflux (GER), though the mechanism underlying this relationship remains unknown. Here we use the validated infant pig model to assess the impacts of simulated GER on preesophageal feeding parameters. We used high-speed videofluoroscopy and electromyography to record bottle-feeding before and following the infusion of a capsaicin-containing solution into the lower esophagus. Sucking parameters were minimally affected by capsaicin exposure, such that genioglossus activity was unchanged and tongue kinematics were largely unaffected. Aspects of the pharyngeal swallow were altered with simulated GER, including increased thyrohyoid muscle activity, increased excursions of the hyoid and thyroid per swallow, decreased swallow frequency, and increased bolus sizes. These results suggest that esophageal afferents can elicit changes in pharyngeal swallowing. In addition, decreased swallowing frequency may be the mechanism by which esophageal pathologies induce oropharyngeal dysphagia. Although recent work indicates that oral or pharyngeal capsaicin may improve dysphagia symptoms, the decreased performance following esophageal capsaicin exposure highlights the importance of designing sensory interventions based upon neurophysiology and the mechanisms underlying disordered feeding. This mechanistic approach requires comprehensive data collection across the entirety of the feeding process, which can be achieved using models such as the infant pig.NEW & NOTEWORTHY Simulated gastroesophageal reflux (GER) in an infant pig model resulted in significant changes in pharyngeal swallowing, which suggests that esophageal afferents are centrally integrated to alter motor outputs to the pharynx. In addition, decreased swallow frequency and increased bolus sizes may be underlying mechanisms by which esophageal pathologies induce oropharyngeal dysphagia. The infant pig model used here allows for a mechanistic approach, which can facilitate the design of intervention strategies based on neurophysiology.

Developmental factors influencing bone strength in precocial mammals: An infant pig model.

Most vertebrates are precocial in locomotion, able to walk and run soon after birth. Precociality requires a bony skeleton of sufficient strength to resist mechanical loading during early locomotor efforts. The aim of this study was to use an animal model-the preterm infant pig-to investigate some of the proximate factors that might determine variation in bone strength in precocial animals. Based on the prior literature, we tested the null predictions that skeletal integrity would be significantly compromised by truncated gestation (i.e., preterm birth) and reduced body mass at birth. We generated a suite of both morphometric measures (tissue mineral density and cross-sectional geometry) and performance-related metrics (ability to resist loading, deformation, and fracture during three-point bending tests) of the appendicular skeleton of preterm and full-term infant pigs. Results showed that very few measures in our ontogenetic infant pig sample significantly varied with either gestation length or birth mass. Overall, our results contribute to a growing body of literature demonstrating the early functional capacity of the precocial infant musculoskeletal system and suggest that bone strength in perinatal precocial mammals may be robust to the factors shown to compromise skeletal integrity in more altricial taxa.

Oropharyngeal capsaicin exposure improves infant feeding performance in an animal model of superior laryngeal nerve damage.

Sensorimotor feedback is critical to safe and effective swallowing. Because of this, sensory interventions have the potential to treat dysphagia. One such treatment may be found in capsaicin, which activates the internal branch of the superior laryngeal nerve (iSLN). The iSLN initiates the pharyngeal swallow, and a more sensitive iSLN should more readily elicit swallowing and improve swallow safety. We explored the neurophysiological mechanism by which capsaicin improves swallow performance using an infant pig model with a unilateral iSLN lesion. Using high-speed videofluoroscopy, we collected oropharyngeal kinematic data while pigs suckled on bottles, before and after applying capsaicin to the posterior tongue and valleculae. We found that capsaicin application decreased maximal bolus sizes, which improved swallow safety. Furthermore, capsaicin improved performance when infant pigs swallowed more moderately sized boluses. However, capsaicin did not change swallow frequency, the number of sucks prior to each swallow, nor total pharyngeal transit time (TPT). Similarly, excursions of the hyoid, thyroid, and posterior tongue were unchanged. TPT and hyoid and thyroid excursions maintained relationships with bolus size post-capsaicin, suggesting that these variables are less sensitive to sensory intervention. The timing and extent of posterior tongue movement were only correlated with bolus size pre-capsaicin, which could imply that capsaicin fundamentally changes in relationships between tongue movements and bolus size. Our results provide insight into the neural control of swallowing and capsaicin's mechanism of action, and suggest that capsaicin may be beneficial in treating acute infant dysphagia.NEW & NOTEWORTHY Chemical sensory interventions alter swallow physiology, which is well-documented in adults but relatively unexplored in infants. Using videofluoroscopy, we found that capsaicin exposure limited infant pigs' bolus sizes to improve swallow performance without changing swallow frequency. Capsaicin increased the likelihood of safe swallowing with more moderately sized boluses and changed relationships between bolus size and tongue movements, which may impact performance. This work highlights the potential role of capsaicin in treating acute infant dysphagia.

Anatomical and physiological variation of the hyoid musculature during swallowing in infant pigs.

The function of a muscle is impacted by its line of action, activity timing and contractile characteristics when active, all of which have the potential to vary within a behavior. One function of the hyoid musculature is to move the hyoid bone during swallowing, yet we have little insight into how their lines of action and contractile characteristics might change during a swallow. We used an infant pig model to quantify the contractile characteristics of four hyoid muscles during a swallow using synchronized electromyography, fluoromicrometry and high-speed biplanar videofluoroscopy. We also estimated muscle line of action during a swallow using contrast-enhanced CT-scanned muscles animated to move with the hyoid bone and found that as the hyoid elevated, the line of action of the muscles attached to it became greater in depression. We also found that muscles acted eccentrically and concentrically, which was correlated with hyoid movement. This work contributes to our understanding of how the musculature powering feeding functions during swallowing.

Swallow Safety is Determined by Bolus Volume During Infant Feeding in an Animal Model.

Feeding difficulties are especially prevalent in preterm infants, although the mechanisms driving these difficulties are poorly understood due to a lack of data on healthy infants. One potential mechanism of dysphagia in adults is correlated with bolus volume. Yet, whether and how bolus volume impacts swallow safety in infant feeding is unknown. A further complication for safe infant swallowing is recurrent laryngeal nerve (RLN) injury due to patent ductus arteriosus surgery, which exacerbates the issues that preterm infants face and can increase the risk of dysphagia. Here, we used a validated animal model feeding freely to test the effect of preterm birth, postnatal maturation and RLN lesion and their interactions on swallow safety. We also tested whether bolus size differed with lesion or birth status, and the relationship between bolus size and swallow safety. We found very little effect of lesion on swallow safety, and preterm infants did not experience more penetration or aspiration than term infants. However, term infants swallowed larger boluses than preterm infants, even after correcting for body size. Bolus size was the primary predictor of penetration or aspiration, with larger boluses being more likely to result in greater degrees of dysphagia irrespective of age or lesion status. These results highlight that penetration and aspiration are likely normal occurrences in infant feeding. Further, when comorbidities, such as RLN lesion or preterm birth are present, limiting bolus size may be an effective means to reduce incidences of penetration and aspiration.

Muscle activity and kinematics show different responses to recurrent laryngeal nerve lesion in mammal swallowing.

Understanding the interactions between neural and musculoskeletal systems is key to identifying mechanisms of functional failure. Mammalian swallowing is a complex, poorly understood motor process. Lesion of the recurrent laryngeal nerve, a sensory and motor nerve of the upper airway, results in airway protection failure (liquid entry into the airway) during swallowing through an unknown mechanism. We examined how muscle and kinematic changes after recurrent laryngeal nerve lesion relate to airway protection in eight infant pigs. We tested two hypotheses: 1) kinematics and muscle function will both change in response to lesion in swallows with and without airway protection failure, and 2) differences in both kinematics and muscle function will predict whether airway protection failure occurs in lesion and intact pigs. We recorded swallowing with high-speed videofluoroscopy and simultaneous electromyography of oropharyngeal muscles pre- and postrecurrent laryngeal nerve lesion. Lesion changed the relationship between airway protection and timing of tongue and hyoid movements. Changes in onset and duration of hyolaryngeal muscles postlesion were less associated with airway protection outcomes. The tongue and hyoid kinematics all predicted airway protection outcomes differently pre- and postlesion. Onset and duration of activity in only one infrahyoid and one suprahyoid muscle showed a change in predictive relationship pre- and postlesion. Kinematics of the tongue and hyoid more directly reflect changes in airway protections pre- and postlesion than muscle activation patterns. Identifying mechanisms of airway protection failure requires specific functional hypotheses that link neural motor outputs to muscle activation to specific movements.NEW & NOTEWORTHY Kinematic and muscle activity patterns of oropharyngeal structures used in swallowing show different patterns of response to lesion of the recurrent laryngeal nerve. Understanding how muscles act on structures to produce behavior is necessary to understand neural control.

Premature birth impacts bolus size and shape through nursing in infant pigs.

BACKGROUND: The formation of a bolus of food is critical for proper feeding function, and there is substantial variation in the size and shape of a bolus prior to a swallow. Preterm infants exhibit decreased abilities to acquire and process food, but how that relates to their bolus size and shape is unknown. Here, we test two hypotheses: (1) that bolus size and shape will differ between term and preterm infants, and (2) bolus size and shape will change longitudinally through development in both term and preterm infants. METHODS: To test these hypotheses, we measured bolus size and shape in preterm and term infant pigs longitudinally through nursing using high-speed videofluoroscopy. RESULTS: Preterm infant pigs swallowed smaller volumes of milk. Although term infants increased the amount of milk per swallow as they aged, preterm infants did not. These changes in bolus volume were also correlated with changes in bolus shape; larger boluses became more elongate as they better filled the available anatomical space of the valleculae. CONCLUSIONS: These results suggest that preterm birth reduces the ability of preterm pigs to increase bolus size as they grow, affecting development in this fragile population. These results highlight that studies on term infant feeding may not translate to preterm infants.

Changes in the coordination between respiration and swallowing from suckling through weaning.

All mammals undergo weaning from milk to solid food. This process requires substantial changes to mammalian oropharyngeal function. The coordination of swallowing and respiration is a crucial component of maintaining airway function throughout feeding and matures over infant development. However, how this coordination is affected by weaning is unknown. In this study, we ask how changes in posture, neural maturation and food properties associated with the weaning affect coordination of respiration and swallowing in a validated infant pig model. We recorded seven piglets feeding before and during the weaning age with liquid milk in a bottle and in a bowl, and solid feed in a bowl. Using videofluoroscopy synchronized with respiration, we found (i) the delay in the onset of inspiration after swallowing does not change with head position, (ii) the delay is different between solid food and bowl drinking at the same age and (iii) the delay increases over time when bottle feeding, suggesting a maturational effect. Significant changes in aerodigestive coordination occur prior to and post-weaning, resulting in distinctive patterns for liquid and solid food. The interplay of maturational timelines of oropharyngeal function at weaning may serve as a locus for behavioural and life-history plasticity.

Effects of Superior Laryngeal Nerve Lesion on Kinematics of Swallowing and Airway Protection in an Infant Pig Model.

The superior laryngeal nerve provides detailed sensory information from the mucosal surfaces of laryngeal structures superior to the vocal folds, including the valleculae. Injury to this nerve results in airway penetration and aspiration. Furthermore, such injuries might have an impact on the function of multiple structures involved in intraoral transport and swallowing due to connections within the brainstem. We sought to determine the effects of a surgical lesion of the superior laryngeal nerve on kinematics of the tongue, hyoid, and epiglottis during swallowing. We implanted radio-opaque markers into five infant pigs under anesthesia. Then we fed milk mixed with contrast agent to the pigs while they were recorded via video fluoroscopy, before and after a surgery to transect the superior laryngeal nerve. We digitized and rated airway protection in 177 swallows. We found that in most animals, swallow duration was shorter after nerve lesion. The hyoid also traveled a shorter distance after lesion. Frequently, individuals reacted differently to the same nerve lesion. We suggest that these differences are due to individual differences in neurological connections. When comparing hyoid kinematics between swallows with successful or failed airway protection, we found more consistency among individuals. This indicates that protecting the airway requires specific sets of kinematic events to occur, regardless of the neurological differences among individuals.

Swallow Safety in Infant Pigs With and Without Recurrent Laryngeal Nerve Lesion.

Aerodigestive coordination is critical for safe feeding in mammals, and failure to do so can result in aspiration. Using an infant pig model, we analyzed the impact of recurrent laryngeal nerve (RLN) lesion on aerodigestive coordination and swallow safety at two time points prior to weaning. We used high-speed videofluoroscopy to record 23 infant pigs longitudinally at two ages (7 days, 17 days) feeding on barium milk. We measured respiration with a plethysmograph and used the Infant Mammalian Penetration-Aspiration Scale (IMPAS) to identify unsafe swallows. We tested for changes in swallow safety longitudinally in control and lesion pigs, and whether there was any interaction between the four different groups. On postnatal day 7, lesioned pigs exhibited differences in the frequency distribution of IMPAS scores relative to control pigs on day 7, and 17 day old lesion and control pigs. There were longitudinal changes in performance following RLN lesion through time, suggesting that the impact of RLN lesion decreases with time, as older lesioned pigs performed similarly to older control pigs. We found minimal differences in the impact of aerodigestive coordination on swallow safety, with shorter delays of inspiration onset reflecting higher rates of penetration in young lesioned pigs. Healthy pigs aspirated at a similar rate to those with an RLN lesion indicating that the occasional occurrence of dysphagia in infants may be a normal behavior.

Reduced Coordination of Hyolaryngeal Elevation and Bolus Movement in a Pig Model of Preterm Infant Swallowing.

Preterm infants often have dysphagia. Because reducing lifetime cumulative exposure to radiation in the context of diagnosis and treatment is a continuing goal of all medical fields which use X-ray imaging, efforts exist to reduce reliance on the gold standard diagnostic tool for dysphagia, VFSS. Alternatives, such as video of external hyolaryngeal movement using video recordings of the anterior surface of the neck, must be evaluated and validated against videofluoroscopy, a task for which non-human animal models are appropriate. In this study, we tested the hypotheses that (1) swallows could be identified equally well from video of external hyolaryngeal movement and bolus movement in videofluoroscopy, and that (2) the two measures would be tightly temporally linked in both term and preterm infant pigs. We recorded 222 swallows in simultaneous and precisely synchronized high-speed videofluoroscopy and high-speed camera films of 4 preterm and 3 term infant pigs drinking milk from a bottle. In term pigs, the two measures consistently identified the same swallows in each image stream. However, in preterm pigs there was a high rate of false positives (~ 10% per feeding sequence) and false negatives (~ 27% per feeding sequence). The timing of hyolaryngeal elevation (external video) and bolus movement (videofluoroscopy) was correlated and consistent in terms pigs, but not in preterm pigs. Magnitude of hyolaryngeal elevation was less in preterm pig swallows than term pig swallows. Absence of epiglottal inversion in preterm pigs was not linked to variation in the timing of the two swallow events. Video of external hyolaryngeal movement, though a reliable swallow indicator in term infant pigs, was unreliable in preterm infant pigs. The coordination of swallowing events differs in preterm and term infant pigs. More research is needed into the distinctive biomechanics of preterm infant pigs.

LVC Timing in Infant Pig Swallowing and the Effect of Safe Swallowing.

Recurrent laryngeal nerve (RLN) injury in neonates, a complication of head and neck surgeries, leads to increased aspiration risk and swallowing dysfunction. The severity of resulting sequelae range from morbidity, such as aspiration pneumonia, to mortality from infection and failure to thrive. The timing of airway protective events including laryngeal vestibule closure (LVC) is implicated in aspiration. We unilaterally transected the RLN in an infant pig model to observe changes in the timing of swallowing kinematics with lesion and aspiration. We recorded swallows using high-speed video-fluoroscopic swallow studies (VFSS) and scored them using the Infant Mammalian Penetration and Aspiration Scale (IMPAS). We hypothesized that changes would occur in swallowing kinematics (1) between RLN lesion and control animals, and (2) among safe swallows (IMPAS 1), penetration swallows (IMPAS 3), and aspiration swallows (IMPAS 7). We observed numerous changes in timing following RLN lesion in safe and unsafe swallows, suggesting pervasive changes in the coordination of oropharyngeal function. The timing of LVC, posterior tongue, and hyoid movements differed between pre- and post-lesion in safe swallows. Posterior tongue kinematics differed for post-lesion swallows with penetration. The timing and duration of LVC and posterior tongue movement differed between aspiration swallows pre- and post-lesion. After lesion, safe swallows and swallows with aspiration differed in timing of LVC, laryngeal vestibule opening, and posterior tongue and hyoid movements. The timing of thyrohyoid muscle activity varied with IMPAS, but not lesion. Further study into the pathophysiology of RLN lesion-induced swallowing dysfunction is important to developing novel therapies.

Maturation of the Coordination Between Respiration and Deglutition with and Without Recurrent Laryngeal Nerve Lesion in an Animal Model.

The timing of the occurrence of a swallow in a respiratory cycle is critical for safe swallowing, and changes with infant development. Infants with damage to the recurrent laryngeal nerve, which receives sensory information from the larynx and supplies the intrinsic muscles of the larynx, experience a significant incidence of dysphagia. Using our validated infant pig model, we determined the interaction between this nerve damage and the coordination between respiration and swallowing during postnatal development. We recorded 23 infant pigs at two ages (neonatal and older, pre-weaning) feeding on milk with barium using simultaneous high-speed videofluoroscopy and measurements of thoracic movement. With a complete linear model, we tested for changes with maturation, and whether these changes are the same in control and lesioned individuals. We found (1) the timing of swallowing and respiration coordination changes with maturation; (2) no overall effect of RLN lesion on the timing of coordination, but (3) a greater magnitude of maturational change occurs with RLN injury. We also determined that animals with no surgical intervention did not differ from animals that had surgery for marker placement and a sham procedure for nerve lesion. The coordination between respiration and swallowing changes in normal, intact individuals to provide increased airway protection prior to weaning. Further, in animals with an RLN lesion, the maturation process has a larger effect. Finally, these results suggest a high level of brainstem sensorimotor interactions with respect to these two functions.

Teaching to varied disciplines and educational levels simultaneously: An innovative approach in a neonatal follow-up clinic.

PURPOSE: Across various health conditions and geographic regions, there remains a dearth of clinicians with the expertise and confidence to identify and manage children with disabilities. At the front line of this crisis are clinician-educators, who are tasked with caring for these unique patients and with training the future workforce. Balancing patient care and clinical instruction responsibilities is particularly challenging when trainees of varied educational levels and specialties report simultaneously. The lack of a standard curriculum further compounds the clinician-educator's teaching demands and threatens the consistency of trainees' learning. Recognizing these challenges in their work in a neonatal follow-up clinic, the authors sought a solution through an established curriculum development process. MATERIALS AND METHODS: A needs assessment survey was conducted to gauge medical trainees' knowledge, skills, and experiences. Applying needs assessment findings, the authors developed a curriculum, which was administered online to several trainee cohorts just prior to rotations in the neonatal follow-up clinic. RESULTS: After completing the curriculum, trainees scored significantly higher on neonatal follow-up knowledge tests. CONCLUSIONS: Providing advance exposure helped to ensure that trainees arrived with comparable basal knowledge, which served as a foundation for more advanced instruction. This curricular approach may be useful across teaching venues, especially those with multi-level or multi-discipline learners.

Surprising absence of heparin in the intestinal mucosa of baby pigs.

Heparin, a member of a family of molecules called glycosaminoglycans, is biosynthesized in mucosal mast cells. This important anticoagulant polysaccharide is primarily produced by extraction of the mast cell-rich intestinal mucosa of hogs. There is concern about our continued ability to supply sufficient heparin to support the worldwide growth of advanced medical procedures from the static population of adult hogs used as food animals. While the intestinal mucosa of adult pigs is rich in anticoagulant heparin (containing a few hundred milligrams per animal), little is known about how the content of heparin changes with animal age. Using sophisticated mass spectral analysis we discovered that heparin was largely absent from the intestinal mucosa of piglets. Moreover, while the related, nonanticoagulant heparan sulfate glycosaminoglycan was present in significant amounts we found little chondroitin sulfate E also associated with mast cells. Histological evaluation of piglet intestinal mucosa showed a very low mast cell content. Respiratory mast cells have been reported in baby pigs suggesting that there was something unique about the piglets used in the current study. These piglets were raised in the relatively clean environment of a university animal facility and treated with antibiotics over their lifetime resulting in a depleted microbiome that greatly reduced the number of mast cells and heparin content of the intestinal mucosal in these animals. Thus, from the current study it remains unclear whether the lack of intestinal mast cell-derived heparin results from the young age of these animals or their exposure to their depleted microbiome.

Animal Models for Dysphagia Studies: What Have We Learnt So Far.

Research using animal models has contributed significantly to realizing the goal of understanding dysfunction and improving the care of patients who suffer from dysphagia. But why should other researchers and the clinicians who see patients day in and day out care about this work? Results from studies of animal models have the potential to change and grow how we think about dysphagia research and practice in general, well beyond applying specific results to human studies. Animal research provides two key contributions to our understanding of dysphagia. The first is a more complete characterization of the physiology of both normal and pathological swallow than is possible in human subjects. The second is suggesting of specific, physiological, targets for development and testing of treatment interventions to improve dysphagia outcomes.

Isolated pharyngeal swallow exists during normal human feeding.

Swallowing is one of the basic activities in humans. The pharynx functions as an airway and a food channel, and a pharyngeal swallow usually occurs after bolus transport from the oral cavity. However, direct fluid infusion through a catheter into the hypopharynx produces a pharyngeal swallow without the oral stage in experimental situations. The purpose of this study was to examine whether a pharyngeal swallow, which is not accompanied by bolus transport, can occur during normal human feeding. Fifty-three healthy volunteers (25-89 years) were recorded, via videofluoroscopic examination of swallowing, during 3 different swallowing trials: command swallow of 10 ml liquid barium, chew-swallow of corned beef, and chew-swallow of a mixture of corned beef and liquid barium. Subsequently each swallow was classified as being either a consecutive pharyngeal swallow (CPS), following transport, or an isolated pharyngeal swallow (IPS), without immediately prior transport. The location of the bolus at swallow initiation was also noted. Of 307 trials, 681 swallows were identified, which included 43 IPS and 638 CPS. IPS only occurred as the first swallow of a trial, but the frequency of IPS did not differ between 28 younger (< 60 years) and 25 older (≥ 60 years) people. Of the three food types, IPS occurred more frequently with the mixed food than with liquid. These results suggest that IPS may represent an airway protective mechanism. In conclusion, IPS occurs in normal swallowing during a daily eating situation. Swallowing is more complex than a simple reflex.

The Physiologic Impact of Unilateral Recurrent Laryngeal Nerve (RLN) Lesion on Infant Oropharyngeal and Esophageal Performance.

Recurrent laryngeal nerve (RLN) injury in neonates, a complication of patent ductus arteriosus corrective surgery, leads to aspiration and swallowing complications. Severity of symptoms and prognosis for recovery are variable. We transected the RLN unilaterally in an infant mammalian animal model to characterize the degree and variability of dysphagia in a controlled experimental setting. We tested the hypotheses that (1) both airway protection and esophageal function would be compromised by lesion, (2) given our design, variability between multiple post-lesion trials would be minimal, and (3) variability among individuals would be minimal. Individuals' swallowing performance was assessed pre- and post-lesion using high speed VFSS. Aspiration was assessed using the Infant Mammalian Penetration-Aspiration Scale (IMPAS). Esophageal function was assessed using two measures devised for this study. Our results indicate that RLN lesion leads to increased frequency of aspiration, and increased esophageal dysfunction, with significant variation in these basic patterns at all levels. On average, aspiration worsened with time post-lesion. Within a single feeding sequence, the distribution of unsafe swallows varied. Individuals changed post-lesion either by increasing average IMPAS score, or by increasing variation in IMPAS score. Unilateral RLN transection resulted in dysphagia with both compromised airway protection and esophageal function. Despite consistent, experimentally controlled injury, significant variation in response to lesion remained. Aspiration following RLN lesion was due to more than unilateral vocal fold paralysis. We suggest that neurological variation underlies this pattern.

The effect of bilateral superior laryngeal nerve lesion on swallowing: a novel method to quantitate aspirated volume and pharyngeal threshold in videofluoroscopy.

The purpose was to determine the effect of bilateral superior laryngeal nerve (SLN) lesion on swallowing threshold volume and the occurrence of aspiration, using a novel measurement technique for videofluoroscopic swallowing studies (VFSS) in infant pigs. We used a novel radiographic phantom to assess volume of the milk containing barium from fluoroscopy. The custom made phantom was firstly calibrated by comparing image intensity of the phantom with known cylinder depths. Secondly, known volume pouches of milk in a pig cadaver were compared to volumes calculated with the phantom. Using these standards, we calculated the volume of milk in the valleculae, esophagus and larynx, for 205 feeding sequences from four infant pigs feeding before and after had bilateral SLN lesions. Swallow safety was assessed using the tested and validated IMPAS (Dysphagia 28(2):178-187, 2013). The log-linear correlation between image intensity values from the phantom filled with barium milk and the known phantom cylinder depths was strong (R (2) > 0.95), as was the calculated volumes of the barium milk pouches. The threshold volume of bolus in the valleculae during feeding was significantly larger after bilateral SLN lesion than in control swallows (p < 0.001). The IMPAS score increased in the lesioned swallows relative to the controls, indicating substantially impaired swallowing (p < 0.001). Bilateral SLN lesion dramatically increased the aspiration incidence and the threshold volume of bolus in valleculae. The use of this phantom permits quantification of the aspirated volume of fluid, allowing for more accurate 3D volume estimation from 2D X-ray in VFSS.

Ontogenetic allometry in the thoracolumbar spine of mammal species with differing gait use.

Vertebrae are serially homologous structures with tight integration through their evolution and development. However, in mammals, the thoracic and lumbar regions are morphologically and functionally differentiated. We test the hypothesis that locomotor specialization is associated with altered post-natal growth patterns in vertebrae from different vertebral regions. We use longitudinal data to examine thoracolumbar growth in two specialized half-bounding (Oryctolagus cuniculus and Chinchilla lanigera) and two non-specialized (Cavia porcellus and Monodelphis domestica) species with similar body sizes. Lateral X-rays of 38 individuals were the source of centrum length, centrum height and intervertebral space length measures for 19-20 thoracolumbar vertebrae. The repeated measurements design included the same individuals soon after birth and again at adult size. Data from columns with different vertebral counts were compared by either summing (length) or averaging (height) within regions, and individual vertebrae were directly compared at the first and last five vertebral positions. Specialized half-bounders had longer lumbar regions than generalists, which was attributable to positively allometric growth of the lumbar centra. Lumbar centrum length was more variable both ontogenetically and interspecifically than the other variables, suggesting heterochrony may be generating lumbar variation. Craniocaudal patterns of centrum growth correlate with expression of regionalizing genes (i.e., Hox).