X-ray Reconstruction Of Moving Morphology Research Articles

The avian vocal system: 3D reconstruction reveals upper vocal tract elongation during head motion.

While birds' impressive singing abilities are made possible by the syrinx, the upper vocal system (i.e., trachea, larynx, and beak) could also play a role in sound filtration. Yet, we still lack a clear understanding of the range of elongation this system can undertake, especially along the trachea. Here, we used biplanar cineradiography and X-ray Reconstruction of Moving Morphology (XROMM) to record 15 species of cadaveric birds from 9 different orders while an operator moved the bird's cadaveric heads in different directions. In all studied species, we found elongation of the trachea to be correlated with neck extension, and significantly greater (ranging from 18% to 48% for the whole motion; and from 1.4% to 15.7% for the singing positions) than previously reported on a live singing bird (3%). This elongation or compression was not always homogeneous along its entire length. Some specimens showed increased lengthening in the rostral part and others in both the rostral and caudal parts of the vocal tract. The diversity of elongation patterns shows that trachea elongation is more complex than previously thought. Since tracheal lengthening affects sound frequencies, our results contribute to our understanding of the mechanisms involved in complex communication signals, one of the amazing traits we share with birds.

Exploring elbow kinematics in the central bearded dragon (Pogona vitticeps) using XROMM: Implications for the role of forearm long-axis rotation in non-avian reptile posture and mobility.

The functional morphology and kinematics of the elbow joint remain relatively understudied in squamates. Previous investigations of lizard elbow morphology and kinematics suggest long-axis rotation (LAR) of the radius and ulna during stance allows the manus to remain pronated during forelimb retraction. Using XROMM (X-ray Reconstruction of Moving Morphology), we explored the range of 3D movements and kinematics of the humerus, radius, and ulna in three adult male Central bearded dragons (Pogona vitticeps) during trackway walking. Our data indicate that the elbow joint of P. vitticeps experiences significant rotations in all three dimensions and that the radius and ulna adduct and rotate laterally on their long axes relative to the elbow joint and to one another during stance. These movements allow the distal ends of the radius and ulna to remain in a configuration necessary for manus pronation. These data support previous inferences that the radius and ulna of lizards move independently at the wrist joint. We suggest that independent LAR of the radius and ulna relative to the elbow joint and to one another may be an ancestral mechanism in lizards and perhaps more broadly across non-avian reptiles.

The functional role of the rabbit digastric muscle during mastication.

Muscle spindle abundance is highly variable in vertebrates, but the functional determinants of this variation are unclear. Recent work has shown that human leg muscles with the lowest abundance of muscle spindles primarily function to lengthen and absorb energy, while muscles with a greater spindle abundance perform active-stretch-shorten cycles with no net work, suggesting that muscle spindle abundance may be underpinned by muscle function. Compared with other mammalian muscles, the digastric muscle contains the lowest abundance of muscle spindles and, therefore, might be expected to generate substantial negative work. However, it is widely hypothesised that as a jaw-opener (anatomically) the digastric muscle would primarily function to depress the jaw, and consequently do positive work. Through a combination of X-ray reconstruction of moving morphology (XROMM), electromyography and fluoromicrometry, we characterised the 3D kinematics of the jaw and digastric muscle during feeding in rabbits. Subsequently, the work loop technique was used to simulate in vivo muscle behaviour in situ, enabling muscle force to be quantified in relation to muscle strain and hence determine the muscle's function during mastication. When functioning on either the working or balancing side, the digastric muscle generates a large amount of positive work during jaw opening, and a large amount of negative work during jaw closing, on average producing a relatively small amount of net negative work. Our data therefore further support the hypothesis that muscle spindle abundance is linked to muscle function; specifically, muscles that absorb a relatively large amount of negative work have a low spindle abundance.

Effects of healthy aging on tongue-jaw kinematics during feeding behavior in rhesus macaques.

Several age-related oral health problems have been associated with neurodegenerative diseases such as Alzheimer's Disease (AD), yet how oromotor dysfunction in healthy aging differ from those found in pathological aging is still unknown. This is partly because changes in the cortical and biomechanical ("neuromechanical") control of oromotor behavior in healthy aging are poorly understood. To this end, we investigated the natural feeding behavior of young and aged rhesus macaques (Macaca mulatta) to understand the age-related differences in tongue and jaw kinematics. We tracked tongue and jaw movements in 3D using high-resolution biplanar videoradiography and X-ray Reconstruction of Moving Morphology (XROMM). Older subjects exhibited a reduced stereotypy in tongue movements during chews and a greater lag in tongue movements relative to jaw movements compared to younger subjects. Overall, our findings reveal age-related changes in tongue and jaw kinematics, which may indicate impaired tongue-jaw coordination. Our results have important implications for the discovery of potential neuromechanical biomarkers for early diagnosis of AD.

Evaluation of a cadaveric wrist motion simulator using marker-based X-ray reconstruction of moving morphology.

Surgical intervention is a common option for the treatment of wrist joint arthritis and traumatic wrist injury. Whether this surgery is arthrodesis or a motion preserving procedure such as arthroplasty, wrist joint biomechanics are inevitably altered. To evaluate effects of surgery on parameters such as range of motion, efficiency and carpal kinematics, repeatable and controlled motion of cadaveric specimens is required. This study describes the development of a device that enables cadaveric wrist motion to be simulated before and after motion preserving surgery in a highly controlled manner. The simulator achieves joint motion through the application of predetermined displacements to the five major tendons of the wrist, and records tendon forces. A pilot experiment using six wrists aimed to evaluate its accuracy and reproducibility. Biplanar X-ray videoradiography (BPVR) and X-Ray Reconstruction of Moving Morphology (XROMM) were used to measure overall wrist angles before and after total wrist arthroplasty. The simulator was able to produce flexion, extension, radioulnar deviation, dart thrower's motion and circumduction within previously reported functional ranges of motion. Pre- and post-surgical wrist angles did not significantly differ. Intra-specimen motion trials were repeatable; root mean square errors between individual trials and average wrist angle and tendon force profiles were below 1° and 2 N respectively. Inter-specimen variation was higher, likely due to anatomical variation and lack of wrist position feedback. In conclusion, combining repeatable intra-specimen cadaveric motion simulation with BPVR and XROMM can be used to determine potential effects of motion preserving surgeries on wrist range of motion and biomechanics.

XROMM Analysis of Feeding Mechanics in Toads: Interactions of the Tongue, Hyoid, and Pectoral Girdle

SynopsisDuring feeding in many terrestrial vertebrates, the tongue acts in concert with the hyoid and pectoral girdle. In frogs, these three elements are interconnected by musculature. While the feeding mechanics of the anuran tongue are well-studied, little is known of how the motions of the tongue relate to the movements of the skeleton or how buccal structures move following closure of the mouth. Although features such as the pectoral girdle and hyoid are not externally visible in frogs, their motions can be tracked in X-ray video. We used XROMM (X-ray Reconstruction of Moving Morphology) techniques to track the 3D movements of the tongue, hyoid apparatus, pectoral girdle, skull, and jaw during the feeding cycle of the cane toad, Rhinella marina. We show how the movements of these elements are integrated during tongue protrusion and prey capture, as well as during prey transport, swallowing, and recovery. Our findings suggest that the hyoid apparatus is important both for prey manipulation and swallowing. The tongue consistently stretches posterior to the skull during swallowing, often more than it stretches during protrusion to reach the prey. Feeding kinematics are similar between individuals, and the kinematics of unsuccessful strikes generally resemble those of successful strikes. Our data also provide a new perspective on the potential role of the pectoral girdle, an element with a predominant locomotor function, during feeding events. This work raises new questions about the evolution of feeding in frogs, as well as how the diversity of pectoral and buccal anatomy observed across anurans may influence feeding kinematics.

Suction feeding of West African lungfish (Protopterus annectens): An XROMM analysis of jaw mechanics, cranial kinesis, and hyoid mobility.

ABSTRACTSuction feeding in fishes is characterized by rapid cranial movements, but extant lungfishes (Sarcopterygii: Dipnoi) exhibit a reduced number and mobility of cranial bones relative to actinopterygian fishes. Despite fusion of cranial elements, lungfishes are proficient at suction feeding, though the impacts of novel cranial morphology and reduced cranial kinesis on feeding remain poorly understood. We used X-ray reconstruction of moving morphology (XROMM) to study the kinematics of seven mobile elements (neurocranium, upper jaw, lower jaw, tongue, ceratohyal, clavicle, and cranial rib) and two muscles (costoclavicular portion of the hypaxialis and rectus cervicis) during the feeding strikes of West African lungfish (Protopterus annectens). We found that feeding by P. annectens on non-evasive prey is relatively slow, with a mean time to peak gape of 273 ms. Lower jaw depression and clavicular rotation were hinge-like, with one degree of freedom, but the ceratohyals rotated in a complex motion involving depression and long-axis rotation. We quantified the relative contributions to oral cavity volume change (RCVC) and found that oral cavity expansion is created primarily by ceratohyal and clavicle motion. P. annectens suction feeds relatively slowly but successfully through muscle shortening of hypaxial and rectus cervicis muscles contributing to hyoid mobility.

Constraining Morphologies of Soft Tissues in Extinct Vertebrates Using Multibody Dynamic Simulations: A Case Study on Articular Cartilage of the Sauropod Dreadnoughtus

Rarity of soft tissue preservation, including of articular cartilage, in the fossil record hinders creation of biologically-realistic mechanical models. Previous studies of articular cartilage in extant taxa have documented important aspects of cartilage shapes and thicknesses, but these insights remain generalized and have yet to see systematic implementation in biomechanical modeling. Herein, we present a new method for modeling joints that allows for testing of hypotheses about articular cartilage morphology in extinct taxa. Our case study examines the left elbow joint of the sauropod dinosaurDreadnoughtus schraniusing articular cartilage reconstructions constrained by extant phylogenetic bracketing (EPB). EPB investigations of alligator and chicken articular cartilage revealed the presence of a spherical anterior projection of cartilage on the distal humerus which articulates with the radius during flexion. Importantly, this shape does not directly mirror the underlying bone. Using multibody dynamic models created in Adams™ withouta priorirestrictions on joint degrees of freedom, we simulated the effects of three alternative cartilage reconstructions based on these EPB findings which differ in mediolateral placement of a cartilage sphere and its anteroposterior thickness, encompassing a range of possibilities for the condition inDreadnoughtus. Bone kinematics and contact area (calculated in Geomagic®) were tracked. Additionally, we modeled the elbow of an alligator and turkey using the same methodology and compared the results to XROMM (X-ray Reconstruction of Moving Morphology) analysis of the same limbs. Each model produced distinct results but were generally similar supporting our modeling methodology. Based on these findings, we predict thatDreadnoughtus, and presumably other extinct archosaurs, had a spherical projection of cartilage on the anterior face of the distal end of the humerus for articulation with the radius. Though many valuable insights have been gained by existing modeling methodologies, we chose a different approach that focused on joint contact surfaces. Moreover, applying our methods within a quantitative hypothesis-testing framework can advance the field of paleobiology by testing hypotheses relating shape and kinematics that are not possible with prescribed joint motions.

Regional Tongue Anatomy and Corresponding Kinematics During Chewing and Drinking in the Pig

Chewing and drinking are fundamental oral behaviors for nutrient ingestion in mammals. They consist of coordinated tongue and jaw movements to ingest, transport, and swallow. However, differences in bolus properties and intraoral management suggest the potential for differences in the movements and deformations performed by the tongue throughout the cycle. Additionally, regional heterogeneity of muscle fascicle orientation is observed throughout the body of the tongue which likely imposes different constraints on the capabilities of different regions to move and deform. The goal of this study is to determine how regional differences in tongue movements and deformations during chewing and drinking in pigs relate to their corresponding muscle fascicle anatomy. Muscle fascicle orientations were quantified using diffusible iodine contrast‐enhanced computed tomography (diceCT) scans relative to the sagittal plane. Cycle parameters and corresponding regional tongue deformations were analyzed during chewing and drinking using X‐ray Reconstruction of Moving Morphology (XROMM) with additional markers implanted in the soft tissues of the tongue (Ohio University Institutional Animal Care and Use Committee: protocol #12‐U‐009). Examination of muscle fascicles highlights clear regional differences in their orientations. XROMM results demonstrate differences in kinematic parameters between chewing and drinking cycles, with drinking cycles exhibiting a smaller amount of jaw movement for all degrees of freedom used during both behaviors: Tx (condylar protraction‐retraction), Rz (jaw pitch), and Ry (jaw yaw). Tongue protraction‐retraction was greater during chewing than drinking. Additionally, during both behaviors, a near‐constant total tongue length was observed, but regional contributions to total tongue length varied significantly between behaviors, with higher magnitudes of length deformations observed during chewing. This work demonstrates that different regions of the tongue do move and deform in different ways, and that they vary between chewing and drinking behaviors. The observed deformations also correspond to the underlying muscle fascicle anatomy as revealed by diceCT.

Skull‐Brain Biomechanics of Headbutting. An XROMM Pilot Study on Goats

In humans, the coup‐contrecoup injury occurs opposite the site of impact when the brain meets the skull, causing intracerebral hemorrhage. It makes up a significant portion of traumatic brain injuries (TBI) and can often lead to neurodegeneration. Bighorn sheep and related bovids are known for their impressive headbutting rituals performed during the rut, the general belief being that these animals come away unharmed from these encounters. However, a recent immunohistochemical study revealed TBI in bighorn sheep and muskoxen in relation to headbutting. With a large gyrencephalic brain, repetitive headbutting, and a pathology pattern resembling early chronic traumatic encephalopathy in humans, bovids present themselves as a significant model for TBI. It is, however, uncertain whether bovids sustain coup‐contrecoup injury due to the oblong shape of their braincase and tight fit to the brain. We aimed to investigate whether cranial and neuroanatomical aspects of the bovid head anatomy contribute to TBI reduction, using goats as a model to measure brain movement independent of the skull upon impact.Four cadaveric goat heads were implanted with radiodense tantalum beads in the skull and brain. A weight‐drop system was used to create an impact within the field of view of a biplanar fluoroscopy system. The heads were CT scanned and the recording was reconstructed as an animated 3D model using x‐ray reconstruction of moving morphology (XROMM). The brain markers were tracked relative to each other to account for soft tissue deformation and marker migration and relative to the skull markers to account for brain movement. We were able to visualize how the brain moves in relation to the skull for the first time in bovids, providing a better understanding of skull‐brain biomechanics.

Suction feeding biomechanics of Polypterus bichir: investigating linkage mechanisms and the contributions of cranial kinesis to oral cavity volume change.

Many fishes use substantial cranial kinesis to rapidly increase buccal cavity volume, pulling prey into the mouth via suction feeding. Living polypterids are a key lineage for understanding the evolution and biomechanics of suction feeding because of their phylogenetic position and unique morphology. Polypterus bichir have fewer mobile cranial elements compared with teleosts [e.g. immobile (pre)maxillae] but successfully generate suction through dorsal, ventral and lateral oral cavity expansion. However, the relative contributions of these motions to suction feeding success have not been quantified. Additionally, extensive body musculature and lack of opercular jaw opening linkages make P. bichir of interest for examining the role of cranial versus axial muscles in driving mandibular depression. Here, we analyzed the kinematics of buccal expansion during suction feeding in P. bichir using X-ray Reconstruction of Moving Morphology (XROMM) and quantified the contributions of skeletal elements to oral cavity volume expansion and prey capture. Mouth gape peaks early in the strike, followed by maximum cleithral and ceratohyal rotations, and finally by opercular and suspensorial abductions, maintaining the anterior-to-posterior movement of water. Using a new method of quantifying bones' relative contributions to volume change (RCVC), we demonstrate that ceratohyal kinematics are the most significant drivers of oral cavity volume change. All measured cranial bone motions, except abduction of the suspensorium, are correlated with prey motion. Lastly, cleithral retraction is largely concurrent with ceratohyal retraction and jaw depression, while the sternohyoideus maintains constant length, suggesting a central role of the axial muscles, cleithrum and ceratohyal in ventral expansion.

Novel biomarkers in healthy aging: Tongue and jaw kinematics as a potential way to further explore Alzheimer’s‐induced biological changes

AbstractBackgroundMany age‐related oral health problems, such as masticatory dysfunction and dysphagia, have been associated with Alzheimer’s Disease (AD). However, how oromotor dysfunctions in healthy aging differ from those found in AD is still largely unknown. Here, we aim to assess changes in tongue and jaw kinematics in healthy aging to understand how these biomarkers may provide early signatures of AD.MethodData were collected from two male monkeys (Macaca mulatta), one young (8 years) and one older (20 years). Using high‐resolution bilateral videoradiography (200 frames/s) and the X‐Ray Reconstruction of Moving Morphology (XROMM) workflow, we quantified the 3D position of tongue and jaw radio‐opaque markers while monkeys engaged in a natural feeding behavior. To evaluate age‐related changes, we analyzed tongue and jaw kinematics relative to specific events in the feeding cycle during 10‐s feeding trials (20‐60 trials). All protocols were approved by the University of Chicago Animal Care and Use Committee and complied with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. ResultDuring chews and swallows, both monkeys exhibited cyclical tongue motions, however, the tongue motions in the older monkey were substantially less stereotyped. The distributions of x‐y‐z positions of the tongue during start/end of chews and swallows were tightly clustered in the young monkey but were more spread out in the older (Fig. 1). In addition, tongue movements in the rostro‐caudal direction lagged behind jaw movements to a greater degree in the older monkey than in the younger (Fig. 2). Indeed, in the young monkey, tongue movements in the rostro‐caudal direction were highly correlated with supero‐inferior jaw movements, with peak correlations occurring at zero lag (Fig. 3).ConclusionThese results show that tongue and jaw kinematics may change with age, particularly with regards to stereotypy of tongue movement and tongue‐jaw coordination. Understanding oromotor changes in healthy aging may serve as diagnostic measures that may be helpful in treating or delaying AD progression and in reducing the severity of debilitating oromotor dysfunctions.

Validation of an Echidna Forelimb Musculoskeletal Model Using XROMM and diceCT.

In evolutionary biomechanics, musculoskeletal computer models of extant and extinct taxa are often used to estimate joint range of motion (ROM) and muscle moment arms (MMAs), two parameters which form the basis of functional inferences. However, relatively few experimental studies have been performed to validate model outputs. Previously, we built a model of the short-beaked echidna (Tachyglossus aculeatus) forelimb using a traditional modelling workflow, and in this study we evaluate its behaviour and outputs using experimental data. The echidna is an unusual animal representing an edge-case for model validation: it uses a unique form of sprawling locomotion, and possesses a suite of derived anatomical features, in addition to other features reminiscent of extinct early relatives of mammals. Here we use diffusible iodine-based contrast-enhanced computed tomography (diceCT) alongside digital and traditional dissection to evaluate muscle attachments, modelled muscle paths, and the effects of model alterations on the MMA outputs. We use X-ray Reconstruction of Moving Morphology (XROMM) to compare ex vivo joint ROM to model estimates based on osteological limits predicted via single-axis rotation, and to calculate experimental MMAs from implanted muscles using a novel geometric method. We also add additional levels of model detail, in the form of muscle architecture, to evaluate how muscle torque might alter the inferences made from MMAs alone, as is typical in evolutionary studies. Our study identifies several key findings that can be applied to future models. 1) A light-touch approach to model building can generate reasonably accurate muscle paths, and small alterations in attachment site seem to have minimal effects on model output. 2) Simultaneous movement through multiple degrees of freedom, including rotations and translation at joints, are necessary to ensure full joint ROM is captured; however, single-axis ROM can provide a reasonable approximation of mobility depending on the modelling objectives. 3) Our geometric method of calculating MMAs is consistent with model-predicted MMAs calculated via partial velocity, and is a potentially useful tool for others to create and validate musculoskeletal models. 4) Inclusion of muscle architecture data can change some functional inferences, but in many cases reinforced conclusions based on MMA alone.

Alligators employ intermetatarsal reconfiguration to modulate plantigrade ground contact.

ABSTRACTFeet must mediate substrate interactions across an animal's entire range of limb poses used in life. Metatarsals, the ‘bones of the sole’, are the dominant pedal skeletal elements for most tetrapods. In plantigrade species that walk on the entirety of their sole, such as living crocodylians, intermetatarsal mobility offers the potential for a continuum of reconfiguration within the foot itself. Alligator hindlimbs are capable of postural extremes from a belly sprawl to a high walk to sharp turns – how does the foot morphology dynamically accommodate these diverse demands? We implemented a hybrid combination of marker-based and markerless X-ray reconstruction of moving morphology (XROMM) to measure 3D metatarsal kinematics in three juvenile American alligators (Alligator mississippiensis) across their locomotor and maneuvering repertoire on a motorized treadmill and flat-surfaced arena. We found that alligators adaptively conformed their metatarsals to the ground, maintaining plantigrade contact throughout a spectrum of limb placements with non-planar feet. Deformation of the metatarsus as a whole occurred through variable abduction (twofold range of spread) and differential metatarsal pitching (45 deg arc of skew). Internally, metatarsals also underwent up to 65 deg of long-axis rotation. Such reorientation, which correlated with skew, was constrained by the overlapping arrangement of the obliquely expanded metatarsal bases. Such a proximally overlapping metatarsal morphology is shared by fossil archosaurs and archosaur relatives. In these extinct taxa, we suggest that intermetatarsal mobility likely played a significant role in maintaining ground contact across plantigrade postural extremes.

Evaluation of True Ankle Motion Following Total Ankle Replacement Utilizing XROMM technology

Category:Ankle ArthritisIntroduction/Purpose:Total ankle replacement (TAR) is common tool used by the foot and ankle specialist to treat end stage ankle arthritis. Current data about ankle motion following TAR is derived from gait analysis utilizing external markers. Utilizing Xray Reconstruction of Moving Morphology (XROMM), which combines 3-D mapping technology with biplanar fluoroscopy in vivo to visualize true skeletal motion, we can evaluate true motion of TAR implants. Current TAR replacement systems are either mobile bearing or fixed bearing. We hypothesized that subjects implanted with a fixed bearing prosthesis would exhibit less tibiotalar rotation and translation than subjects implanted with a mobile bearing prosthesis.Methods:Six subjects with total ankle replacement at least one-year post implantation gave informed consent before participating (IRB #H16496). Three subjects with a mobile bearing prosthesis with an average age 63.3+-11.1 yrs were compared to three matched subjects with a fixed bearing prosthesis with an average age of 64.7+-1.5 yrs. Utilizing 3D slicer software, lower body CT scans for each subject were evaluated to create 3D models of the foot and ankle bones and implant components. All subjects walked for several trials at a self-selected pace along a walkway while their foot and ankle motions were captured by a high-speed biplanar fluoroscopic x-ray motion analysis (XMA) system. The 3D models were combined with the x-ray images within a 3D animation platform and rotoscoped to resolve accurate kinematic motions at the tibiotalar joint during stance phase of gait. We examined for differences between the two groups using a two-sample t-test (p<0.05).Results:Subjects with a mobile-bearing prosthesis demonstrated mean ROM’s of 7.4+-1.1°, 5.3+-2.3° and 7.1+-4.3° for dorsiflexion/plantarflexion, inversion/eversion, and internal/external rotation, respectively. Subjects with a fixed bearing ankle prosthesis did not exhibit significantly different mean ROM’s for dorsiflexion/plantarflexion (9.1+-4.0°, p=0.35), inversion/eversion (4.4+-2.1°, p=0.42), and internal/external rotation (9.0+-3.4°, p=0.35), respectively. Subjects with a fixed bearing prosthesis displayed significantly more translation along the anteroposterior (3.6+-1.2mm, p<0.01) and mediolateral (2.2+-0.7mm, p<0.01) axes compared to the mobile bearing prosthesis (1.8+-1.2mm and 1.3+-0.8mm, respectively).Conclusion:Our preliminary results indicate that mobile and fixed bearing prosthesis provides similar angular motion at the tibiotalar joint, however, the fixed bearing prosthesis exhibits greater translational motion during walking. Further, there is the same amount of internal and external translation with both component designs. The implications of this work on success or failure of current implant designs is beyond the scope of this study but this work will provide the basis for future studies to help determine optimal future total ankle replacement designs.

Effects of food properties on chewing in pigs: Flexibility and stereotypy of jaw movements in a mammalian omnivore

Chewing is a rhythmic oral behavior that requires constant modifications of jaw movements in response to changes in food properties. The food-specific kinematic response is dependent on the potential for kinematic flexibility allowed by morphology and modulation of motor control. This study investigates the effects of food toughness and stiffness on the amplitude and variability of jaw movements during chewing in a typical omnivorous mammalian model (pigs). Jaw movements were reconstructed using X-ray Reconstruction Of Moving Morphology (XROMM) and kinematic data associated with the amplitude of jaw pitch (opening-closing) and jaw yaw (mediolateral rotation) were extracted for each cycle. Between-food differences were tested for the amplitude of jaw movements during each phase of the gape cycle, as well as in their respective within-food variability, or stereotypy, as indicated by coefficients of variation. With increasing toughness, jaw pitch amplitude is decreased during fast close, larger and more stereotyped during slow close, smaller but more variable during slow open, and more variable during fast open. In addition, when chewing on tougher foods, the amplitude of jaw yaw during slow close only increases in a subset of individuals, but all become less variable (i.e., more stereotyped). In contrast, increasing food stiffness has no effect on the amplitude or the variability of jaw pitch, whereas jaw yaw increases significantly in the majority of individuals studied. Our data demonstrate that food stiffness and toughness both play a role in modulating gape cycle dynamics by altering the trajectory of jaw movements, especially during the slow-close phase and tooth-food-tooth contact, albeit differently. This highlights how a generalist oral morphology such as that of pigs (e.g., bunodont teeth lacking precise occlusion, permissive temporomandibular joint allowing extensive condylar displacements in 3 dimensions) enables organisms to not only adjust chewing movements in their amplitude, but also in their variability.

Integrating XMALab and DeepLabCut for high-throughput XROMM.

Marker tracking is a major bottleneck in studies involving X-ray reconstruction of moving morphology (XROMM). Here, we tested whether DeepLabCut, a new deep learning package built for markerless tracking, could be applied to videoradiographic data to improve data processing throughput. Our novel workflow integrates XMALab, the existing XROMM marker tracking software, and DeepLabCut while retaining each program's utility. XMALab is used for generating training datasets, error correction and 3D reconstruction, whereas the majority of marker tracking is transferred to DeepLabCut for automatic batch processing. In the two case studies that involved an in vivo behavior, our workflow achieved a 6 to 13-fold increase in data throughput. In the third case study, which involved an acyclic, post-mortem manipulation, DeepLabCut struggled to generalize to the range of novel poses and did not surpass the throughput of XMALab alone. Deployed in the proper context, this new workflow facilitates large scale XROMM studies that were previously precluded by software constraints.

Contrast-enhanced XROMM reveals in vivo soft tissue interactions in the hip of Alligator mississippiensis.

Extant archosaurs exhibit highly divergent articular soft tissue anatomies between avian and crocodilian lineages. However, the general lack of understanding of the dynamic interactions among archosaur joint soft tissues has hampered further inferences about the function and evolution of these joints. Here we use contrast-enhanced computed tomography to generate 3D surface models of the pelvis, femora, and hip joint soft tissues in an extant archosaur, the American alligator. The hip joints were then animated using marker-based X-Ray Reconstruction of Moving Morphology (XROMM) to visualize soft tissue articulation during forward terrestrial locomotion. We found that the anatomical femoral head of the alligator travels beyond the cranial extent of the bony acetabulum and does not act as a central pivot, as has been suggested for some extinct archosaurs. Additionally, the fibrocartilaginous surfaces of the alligator's antitrochanter and femoral neck remain engaged during hip flexion and extension, similar to the articulation between homologous structures in birds. Moreover, the femoral insertion of the ligamentum capitis moves dorsoventrally against the membrane-bound portion of the medial acetabular wall, suggesting that the inner acetabular foramen constrains the excursion of this ligament as it undergoes cyclical stretching during the step cycle. Finally, the articular surface of the femoral cartilage model interpenetrates with those of the acetabular labrum and antitrochanter menisci; we interpret such interpenetration as evidence of compressive deformation of the labrum and of sliding movement of the menisci. Our data illustrate the utility of XROMM for studying in vivo articular soft tissue interactions. These results also allow us to propose functional hypotheses for crocodilian hip joint soft tissues, expanding our knowledge of vertebrate connective tissue biology and the role of joint soft tissues in locomotor behavior.

How moles walk; it's all thumbs.

A recurring theme in the evolution of tetrapods is the shift from sprawling posture with laterally orientated limbs to erect posture with the limbs extending below the body. However, in order to invade particular locomotor niches, some tetrapods secondarily evolved a sprawled posture. This includes moles, some of the most specialized digging tetrapods. Although their forelimb anatomy and posture facilitates burrowing, moles also walk long distances to forage for and transport food. Here, we use X-ray Reconstruction Of Moving Morphology (XROMM) to determine if the mole humerus rotates around its long axis during walking, as it does when moles burrow and echidnas walk, or alternatively protracts and retracts at the shoulder in the horizontal plane as seen in sprawling reptiles. Our results reject both hypotheses and demonstrate that forelimb kinematics during mole walking are unusual among those described for tetrapods. The humerus is retracted and protracted in the parasagittal plane above, rather than below the shoulder joint and the 'false thumb', a sesamoid bone (os falciforme), supports body weight during the stance phase, which is relatively short. Our findings broaden our understanding of the diversity of tetrapod limb posture and locomotor evolution, demonstrate the importance of X-ray-based techniques for revealing hidden kinematics and highlight the importance of examining locomotor function at the level of individual joint mobility.

Acoustic Emissions as a Non-invasive Biomarker of the Structural Health of the Knee.

The longitudinal assessment of joint health is a long-standing issue in the management of musculoskeletal injuries. The acoustic emissions (AEs) produced by joint articulation could serve as a biomarker for joint health assessment, but their use has been limited by a lack of mechanistic understanding of their creation. In this paper, we investigate that mechanism using an injury model in human lower-limb cadavers, and relate AEs to joint kinematics. Using our custom joint sound recording system, we recorded the AEs from nine cadaver legs in four stages: at baseline, after a sham surgery, after a meniscus tear, and post-meniscectomy. We compare the resulting AEs using their b-values. We then compare joint anatomy/kinematics to the AEs using the X-ray reconstruction of moving morphology (XROMM) technique. After the meniscus tear the number and amplitude of the AE peaks greatly increased from baseline and sham (b-value = 1.33 ± 0.15; p < 0.05). The XROMM analysis showed a close correlation between the minimal inter-joint distances (0.251 ± 0.082cm during extension, 0.265 ± .003 during flexion, at 145°) and a large increase in the AEs. This work provides key insight into the nature of joint AEs, and details a novel technique and analysis for recording and interpreting these biosignals.