Morphologists have historically had to rely on destructive procedures to visualize the three‐dimensional (3‐D) anatomy of animals. More recently, however, non‐destructive techniques have come to the forefront. These include X‐ray computed tomography (CT), which has been used most commonly to examine the mineralized, hard‐tissue anatomy of living and fossil metazoans. One relatively new and potentially transformative aspect of current CT‐based research is the use of chemical agents to render visible, and differentiate between, soft‐tissue structures in X‐ray images. Specifically, iodine has emerged as one of the most widely used of these contrast agents among animal morphologists due to its ease of handling, cost effectiveness, and differential affinities for major types of soft tissues. The rapid adoption of iodine‐based contrast agents has resulted in a proliferation of distinct specimen preparations and scanning parameter choices, as well as an increasing variety of imaging hardware and software preferences. Here we provide a critical review of the recent contributions to iodine‐based, contrast‐enhanced CT research to enable researchers just beginning to employ contrast enhancement to make sense of this complex new landscape of methodologies. We provide a detailed summary of recent case studies, assess factors that govern success at each step of the specimen storage, preparation, and imaging processes, and make recommendations for standardizing both techniques and reporting practices. Finally, we discuss potential cutting‐edge applications of diffusible iodine‐based contrast‐enhanced computed tomography (diceCT) and the issues that must still be overcome to facilitate the broader adoption of diceCT going forward.

Pre-clinical anatomy curricula must provide medical students with the knowledge needed in a variety of medical and surgical specialties. But do physicians within specialties agree about what anatomical knowledge is most important in their practices? And, what is the common core of anatomical knowledge deemed essential by physicians in different specialties? Answers to these questions would be useful in designing pre-clinical anatomy courses. The primary aim of this study was to assess the importance of a human gross anatomy course by soliciting the opinions of physicians from a range of specialties. We surveyed 93 physicians to determine the importance of specific anatomical topics in their own practices. Their responses were analyzed to assess variation in intra- and inter-departmental attitudes toward the importance of anatomy. Nearly all of the topics taught in the course were deemed important by the clinicians as a group, but respondents showed little agreement on the rank order of importance of anatomical topics. Overall, only medical imaging received high importance by nearly all respondents, and lower importance was attached to embryology and lymphatic anatomy. Our survey data, however, also suggested distinct hierarchies in the importance assigned to anatomical topics within specialties. Given that physicians view the importance of anatomy differently, we suggest that students revisit anatomy through a vertically integrated curriculum tailored to provide specialty-specific anatomical training to advanced students based on their areas of clinical interest. Integration of medical imaging into pre-clinical anatomy courses, already underway in many medical schools, is of high clinical relevance.

The tradeoff between force and velocity in skeletal muscle is a fundamental constraint on vertebrate musculoskeletal design (form:function relationships). Understanding how and why different lineages address this biomechanical problem is an important goal of vertebrate musculoskeletal functional morphology. Our ability to answer questions about the different solutions to this tradeoff has been significantly improved by recent advances in techniques for quantifying musculoskeletal morphology and movement. Herein, we have three objectives: (1) review the morphological and physiological parameters that affect muscle function and how these parameters interact; (2) discuss the necessity of integrating morphological and physiological lines of evidence to understand muscle function and the new, high resolution imaging technologies that do so; and (3) present a method that integrates high spatiotemporal resolution motion capture (XROMM, including its corollary fluoromicrometry), high resolution soft tissue imaging (diceCT), and electromyography to study musculoskeletal dynamics in vivo. The method is demonstrated using a case study of in vivo primate hyolingual biomechanics during chewing and swallowing. A sensitivity analysis demonstrates that small deviations in reconstructed hyoid muscle attachment site location introduce an average error of 13.2% to in vivo muscle kinematics. The observed hyoid and muscle kinematics suggest that hyoid elevation is produced by multiple muscles and that fascicle rotation and tendon strain decouple fascicle strain from hyoid movement and whole muscle length. Lastly, we highlight current limitations of these techniques, some of which will likely soon be overcome through methodological improvements, and some of which are inherent. Anat Rec, 301:378-406, 2018. © 2018 Wiley Periodicals, Inc.

Background Cerebral cavernous malformations (CCMs) are hemorrhagic brain lesions, where murine models allow major mechanistic discoveries, ushering genetic manipulations and preclinical assessment of therapies. Histology for lesion counting and morphometry is essential yet tedious and time consuming. We herein describe the application and validations of X-ray micro-computed tomography (micro-CT), a nondestructive technique allowing three-dimensional CCM lesion count and volumetric measurements, in transgenic murine brains. New Method We hereby describe a new contrast soaking technique not previously applied to murine models of CCM disease. Volumetric segmentation and image processing paradigm allowed for histologic correlations and quantitative validations not previously reported with the micro-CT technique in brain vascular disease. Results Twenty-two hyper-dense areas on micro-CT images, identified as CCM lesions, were matched by histology. The inter-rater reliability analysis showed strong consistency in the CCM lesion identification and staging (K=0.89, p<0.0001) between the two techniques. Micro-CT revealed a 29% greater CCM lesion detection efficiency, and 80% improved time efficiency. Comparison with Existing Method Serial integrated lesional area by histology showed a strong positive correlation with micro-CT estimated volume (r2= 0.84, p<0.0001). Conclusions Micro-CT allows high throughput assessment of lesion count and volume in pre-clinical murine models of CCM. This approach complements histology with improved accuracy and efficiency, and can be applied for lesion burden assessment in other brain diseases.

Studies of mechanisms of feeding behavior are important in a society where aging- and disease-related feeding disorders are increasingly prevalent. It is important to evaluate the clinical relevance of animal models of the disease and the control. Our present study quantifies macaque hyolingual and jaw kinematics around swallowing cycles to determine the extent to which macaque swallowing resembles that of humans. One female and one male adult Macaca mulatta were trained to feed in a primate chair. Videofluoroscopy was used to record kinematics in a sagittal view during natural feeding on solid food, and the kinematics of the hyoid bone, thyroid cartilage, mandibular jaw, and anterior-, middle-, and posterior-tongue. Jaw gape cycles were defined by consecutive maximum gapes, and the kinematics of the swallow cycles were compared with those of the two consecutive non-swallow cycles preceding and succeeding the swallow cycles. Although there are size differences between macaques and humans, and macaques have shorter durations of jaw gape cycles and hyoid and thyroid upward movements, there are several important similarities between our macaque data and human data reported in the literature: (1) The durations of jaw gape cycles during swallow cycles are longer than those of non-swallow cycles as a result of an increased duration of the jaw-opening phase; (2) Hyoid and thyroid upward movement is linked with a posterior tongue movement and is faster during swallow than non-swallow cycles; (3) Tongue elevation propagates from anterior to posterior during swallow and non-swallow cycles. These findings suggest that macaques can be a useful experimental model for human swallowing studies.

During primate swallowing, tongue base retraction (tBR) drives the food bolus across the oropharynx towards the esophagus and flips the epiglottis over the laryngeal inlet, protecting against penetration and aspiration of food into the airway. Despite the importance of tBR for swallowing performance, the mechanics of tBR are poorly understood. Using biplanar videoradiography (XRoMM) of four macaque monkeys, we tested the extrinsic muscle shortening hypothesis, which posits that shortening of the hyoglossus and styloglossus muscles pulls the tongue base posteriorly, and the muscular hydrostat or intrinsic tongue muscle hypothesis, which suggests that, because the tongue is composed of incompressible fluid, intrinsic muscle shortening increases tongue length and displaces the tongue base posteriorly. our data falsify these hypotheses. instead we suggest a novel hydraulic mechanism of tBR: shortening and rotation of suprahyoid muscles compresses the tongue between the hard palate, hyoid and mouth floor, squeezing the midline tongue base and food bolus back into the oropharynx. our hydraulic mechanism is consistent with available data on human tongue swallowing kinematics. Rehabilitation for poor tongue base retraction might benefit from including suprahyoid muscle exercises during treatment. Retraction of the tongue base against the posterior pharyngeal wall is vital a part of swallowing in mammals, including humans 1-4 but how this retraction happens is not well understood. Two mechanisms have been hypothesized. The extrinsic muscle shortening hypothesis posits that contraction of the hyoglossus and styloglossus muscles pulls the tongue base posteriorly 4-9. In support of this hypothesis, the lines of action of the styloglossus and hyoglossus muscles both have posteriorly-oriented components and these muscles are active during swallowing in many mammals, including humans 9-13. The muscular hydrostat or intrinsic tongue muscle hypothesis suggests that, because the tongue is largely composed of incompressible fluid, reduction in tongue base width due to contraction of the transversely oriented intrinsic tongue muscles must be associated with increases in posterior tongue length, and hence tongue base retraction (TBR) 6,14,15. In support of this mechanism, dynamic magnetic resonance imaging (MRI) of humans during swallowing reveals increases in posterior tongue length and depth that are hypothesized to be caused by decreases in tongue width 6-8. Despite widespread acceptance of these two hypotheses, both mechanisms rest on untested assumptions. Contraction (shortening) of styloglossus, palatoglossus, and hyoglossus muscles during swallowing, which is central to the extrinsic muscle shortening hypothesis, has not yet been demonstrated. Although these muscles are active during swallowing 9,10,16 , muscle activity need not be associated with muscle shortening: claims of muscle function require measurements of both muscle activity and muscle velocity 17-20. Moreover, styloglossus, palatoglossus and hyoglossus muscles inse...

Three-dimensional (3D) tongue movements are central to performance of feeding functions by mammals and other tetrapods, but 3D tongue kinematics during feeding are poorly understood. Tongue kinematics were recorded during grape chewing by macaque primates using biplanar videoradiography. Complex shape changes in the tongue during chewing are dominated by a combination of flexion in the tongue's sagittal planes and roll about its long axis. As hypothesized for humans, in macaques during tongue retraction, the middle (molar region) of the tongue rolls to the chewing (working) side simultaneous with sagittal flexion, while the tongue tip flexes to the other (balancing) side. Twisting and flexion reach their maxima early in the fast close phase of chewing cycles, positioning the food bolus between the approaching teeth prior to the power stroke. Although 3D tongue kinematics undoubtedly vary with food type, the mechanical role of this movement—placing the food bolus on the post-canine teeth for breakdown—is likely to be a powerful constraint on tongue kinematics during this phase of the chewing cycle. The muscular drivers of these movements are likely to include a combination of intrinsic and extrinsic tongue muscles.

The organization of tissues in appendages often affects their mechanical properties and function. In the fish family Labridae, swimming behavior is associated with pectoral fin flexural stiffness and morphology, where fins range on a continuum from stiff to relatively flexible fins. Across this diversity, pectoral fin flexural stiffness decreases exponentially along the length of any given fin ray, and ray stiffness decreases along the chord of the fin from the leading to trailing edge. In this study, we examine the morphological properties of fin rays, including the effective modulus in bending (E), second moment of area (I), segmentation, and branching patterns, and their impact on fin ray stiffness. We quantify intrinsic pectoral fin ray stiffness in similarly sized fins of two closely related species that employ fins of divergent mechanics, the flapping Gomphosus varius and the rowing Halichoeres bivittatus. While segmentation patterns and E were similar between species, measurements of I and the number of fin ray branch nodes were greater in G. varius than in H. bivittatus. A multiple regression model found that of these variables, I was always significantly correlated with fin ray flexural stiffness and that variation in I always explained the majority of the variation in flexural stiffness. Thus, while most of the morphological variables quantified in this study correlate with fin ray flexural stiffness, second moment of area is the greatest factor contributing to variation in flexural stiffness. Further, interspecific variation in fin ray branching pattern could be used as a means of tuning the effective stiffness of the fin webbing to differences in swimming behavior and hydrodynamics. The comparison of these results to other systems begins to unveil fundamental morphological features of biological beams and yields insight into the role of mechanical properties in fin deformation for aquatic locomotion.