Anne M. Burrows

Finite element analysis of performance in the skulls of marmosets and tamarins

Reliance on plant exudates is a relatively rare dietary specialization among mammals. One well‐studied example of closely related exudate feeders is the New World marmosets and tamarins. Whereas marmosets actively gouge tree bark with their incisors to stimulate the flow of sap, tamarins are opportunistic exudate feeders that do not gouge bark. Several studies of the dentaries and jaw adductors indicate that marmosets exhibit specializations for increased gape at the expense of bite force. Few studies, however, have looked to the cranium of marmosets for evidence of functional specializations. Using 3D finite element models of the marmoset Callithrix jacchus and the tamarin Saguinus fuscicollis, we investigated the performance of the cranium under loading regimes that mimicked unilateral molar biting and bark‐gouging. We investigated three measures of performance: the efficiency with which muscle force is transferred to bite force, the extent to which the models are stressed (a predictor of failure), and the work expended by muscles as they deform the skull (total strain energy). We found that during molar biting the two models exhibited similar levels of performance, though the Saguinus model had slightly higher mechanical efficiency, a slightly lower state of stress, and expended more energy on deformation. In contrast, under the bark‐gouging load, Callithrix exhibited much higher mechanical efficiency than Saguinas, but did so at the expense of more work and higher levels of von Mises stress. This analysis illustrates that differences in the shapes of the skulls of Callithrix and Saguinus confer differences in performance. Whether these aspects of performance are targets of selection awaits broader comparative analyses.

EquiFACS: The Equine Facial Action Coding System

Although previous studies of horses have investigated their facial expressions in specific contexts, e.g. pain, until now there has been no methodology available that documents all the possible facial movements of the horse and provides a way to record all potential facial configurations. This is essential for an objective description of horse facial expressions across a range of contexts that reflect different emotional states. Facial Action Coding Systems (FACS) provide a systematic methodology of identifying and coding facial expressions on the basis of underlying facial musculature and muscle movement. FACS are anatomically based and document all possible facial movements rather than a configuration of movements associated with a particular situation. Consequently, FACS can be applied as a tool for a wide range of research questions. We developed FACS for the domestic horse (Equus caballus) through anatomical investigation of the underlying musculature and subsequent analysis of naturally occurring behaviour captured on high quality video. Discrete facial movements were identified and described in terms of the underlying muscle contractions, in correspondence with previous FACS systems. The reliability of others to be able to learn this system (EquiFACS) and consistently code behavioural sequences was high—and this included people with no previous experience of horses. A wide range of facial movements were identified, including many that are also seen in primates and other domestic animals (dogs and cats). EquiFACS provides a method that can now be used to document the facial movements associated with different social contexts and thus to address questions relevant to understanding social cognition and comparative psychology, as well as informing current veterinary and animal welfare practices.

Testing the validity of metacarpal use in sex assessment of human skeletal remains.

To assess the potential of employing metacarpals in assessing sex of human skeletal remains, previous investigators have generated regression equations (Scheuer & Elkington, 1995) and linear discriminant functions (Falsetti, 1995; Stojanowski, 1999) based upon measurements from metacarpals. Results have varied in overall accuracy and which metacarpal produces the greatest accuracy. Using a contemporary sample, this study seeks to evaluate the validity of using metacarpals to assign sex by testing methodologies of previous studies. Measurements defined by previous authors were repeated on metacarpals from 23 adult cadavers and data were subjected to regression equations and linear discriminant analysis according to previous methodologies. Accuracy in sex determination from methods of Scheuer & Elkington (1993) and Falsetti (1995) were lower than originally reported while accuracy from methods of Stojanowski (1999) were higher than previously reported. These results suggest that the use of metacarpals in sex determination may be limited and should be applied cautiously.

Internal Calvarial Bone Distraction in Rabbits with Delayed-Onset Coronal Suture Synostosis

&NA; Recent studies have identified a subpopulation of craniosynostotic individuals who exhibit progressive or delayed‐onset synostosis and mild craniofacial growth abnormalities. These individuals may be good candidates for nonextirpation, distraction osteogenesis therapy. The present study was designed to test this hypothesis by using internal calvarial bone distraction in a rabbit model with familial delayed‐onset craniosynostosis. Data were collected from 159 rabbits: 71 normal controls, 72 with delayed‐onset coronal suture synostosis, 8 with delayed‐onset coronal suture synostosis and coronal suturectomy, and 8 with delayed‐onset coronal suture synostosis and distraction. At 10 days of age, all rabbits had amalgam markers placed on both sides of the frontonasal, coronal, and anterior lambdoidal sutures. At 25 days of age, correction was accomplished through either a 5‐mm‐wide suturectomy or distraction osteogenesis. An internal distraction appliance was fixed to the frontal and parietal bones and percutaneously and intermittently activated at an average of 0.10 mm/day for 42 days (4.11 mm total). Serial radiographs were taken at 10, 25, 42, and 84 days of age. Results revealed that rabbits with delayed‐onset synostosis had significantly (p < 0.01) reduced coronal suture growth rates (0.04 mm/day) compared with the other three groups (0.07 mm/day). Rabbits with suturectomy and rabbits with distraction showed similar coronal suture responses. However, from 42 to 84 days of age, rabbits with distraction showed reduced growth at the vault sutures and abnormal growth patterns in cranial vault width, cranial vault shape, and cranial base angulation compared with the other three groups. Results demonstrated that, although the normal coronal suture growth rate was maintained in rabbits with delayed‐onset synostosis using intermittent distraction osteogenesis, normal adult craniofacial structure was not achieved. Such anomalous growth was probably a result of altered growth vectors and compressive forces at adjacent sutures during distraction. These findings suggest that distraction osteogenesis without corticotomy may be a treatment alternative in individuals with progressive, delayed‐onset synostosis, but that internal appliances that generate low‐level, continuous distractive forces should be investigated and developed. (Plast. Reconstr. Surg. 102: 1109, 1998.)

Blocking Bone Morphogenetic Protein Function Using In Vivo Noggin Therapy Does Not Rescue Premature Suture Fusion in Rabbits with Delayed-Onset Craniosynostosis

Background: Craniosynostosis is defined as the premature fusion of one or more cranial sutures. Bone morphogenetic proteins (BMPs), regulators of ossification, have been implicated in premature suture fusion. Noggin, an extracellular BMP inhibitor, has been shown experimentally to inhibit resynostosis following surgery. The present study was designed to test the hypothesis that BMP inhibition using noggin therapy may rescue sutures destined to fuse by inhibiting initial ossification. Methods: Twenty-six, 10-day old rabbits with familial, delayed-onset, coronal suture synostosis were randomly divided into three groups: (1) the sham surgical control group, (2) the bovine serum albumin–treated group [10 &mgr;g/suture (protein/vehicle controls)], and (3) the noggin therapy group (10 &mgr;g/suture; experimental group). Sutural growth was monitored by radiopaque markers implanted at 10 days of age. At 25 days, the bovine serum albumin or noggin was combined with a slow-resorbing collagen vehicle and injected subperiosteally above the coronal suture. Somatic and sutural growth data were collected at 10, 25, 42, and 84 days of age. Coronal sutures were harvested at 84 days to histologically assess fusion. Results: Results showed no significant (p > 0.05) differences in suture separation at any age. Suture fusion assessed by histomorphology did not differ among the three groups. Although previous data showed noggin to inhibit postoperative resynostosis in this craniosynostotic rabbit model, here there was no effect on initial suture fusion. Conclusion: These results suggest that in this rabbit model of craniosynostosis, BMPs do not play a role in the pathogenesis of craniosynostosis and only play a role in postoperative bony wound healing.

Eye Size and Set in Small‐Bodied Fossil Primates: A Three‐Dimensional Method

We introduce a new method to geometrically reconstruct eye volume and placement in small‐bodied primates based on the three‐dimensional contour of the intraorbital surface. We validate it using seven species of living primates, with dry skulls and wet dissections, and test its application on seven species of Paleogene fossils of interest. The method performs well even when the orbit is damaged and incomplete, lacking the postorbital bar and represented only by the orbital floor. Eye volume is an important quantity for anatomic and metabolic reasons, which due to differences in eye set, or position within (or outside) the bony orbit, can be underestimated in living and fossil forms when calculated from aperture diameter. Our Ectopic Index quantifies how much the globes volume protrudes anteriorly from the aperture. Lemur, Notharctus and Rooneyia resemble anthropoids, with deeply recessed eyes protruding 11%–13%. Galago and Tarsius are the other extreme, at 47%–56%. We argue that a laterally oriented aperture has little to do with line‐of‐sight in euprimates, as large ectopic eyes can position the cornea to enable a directly forward viewing axis, and soft tissue positions the eyes facing forward in megachiropteran bats, which have unenclosed, open eye sockets. The size and set of virtual eyes reconstructed from 3D cranial models confirm that eyes were large to hypertrophic in Hemiacodon, Necrolemur, Microchoerus, Pseudoloris and Shoshonius, but eye size in Rooneyia may have been underestimated by measuring the aperture, as in Aotus. Anat Rec, 299:1671–1689, 2016.

Relaxin Does Not Rescue Coronal Suture Fusion in Craniosynostotic Rabbits

Objectives Craniosynostosis affects 1 in 2000 to 3000 live births and may result in craniofacial and neural growth disturbances. Histological data have shown that thick collagenous bundles are present in the sutural ligament, which may tether the osteogenic fronts, resulting in premature fusion. The hormone relaxin has been shown to disrupt collagen fiber organization, possibly preventing craniosynostosis by relaxing the sutural ligament and allowing osteogenic fronts to separate normally and stay patent. This study tested this hypothesis with a rabbit model of delayed-onset coronal suture synostosis. Methods A total of 18 New Zealand White rabbits with craniosynostosis were randomly assigned to one of three groups: sham control, protein control (BSA), relaxin treatment. After initial diagnosis, sham surgery, BSA, or relaxin was delivered to the fusing coronal suture in a slow-release (56-day) collagen vehicle. Longitudinal radiographs and body weights were collected at 10, 25, 42, and 84 days of age, and sutures were harvested for histology. Results Relaxin-treated animals had more disorganized intrasuture content than control groups. These specimens also appeared to have relatively wider sutures ectocranially. There were no significant differences in relaxin-treated animals for all craniofacial growth measures, or suture separation compared with controls. Conclusions These data do not support our initial hypothesis that the use of relaxin may rescue sutures destined to undergo premature suture fusion. These findings suggest that collagen fiber arrangement may not be important for suture fusion. This protein therapy would not be clinically useful for craniosynostosis.

Transforming growth factor-β3 therapy delays Postoperative reossification and improves craniofacial growth in craniosynostotic rabbits

Postoperative reossification is a common clinical correlate following surgery. It has been suggested that an underexpression of transforming growth factor-β3 (TGF-β3) may be related to craniosynostosis and postoperative reossification. Adding TGF-β3 may delay reossification and improve postoperative growth. The present study was designed to test this hypothesis. Thirty 10-day-old New Zealand white rabbits with hereditary coronal suture synostosis were divided into three groups: (1) suturectomy controls (n = 14), (2) suturectomy treated with bovine serum albumin (n = 8), and (3) suturectomy treated with TGF-β3 protein (n = 8). At 10 days of age, a 3-mm × 15-mm coronal suturectomy was performed, and serial three-dimensional (3D) computed tomography (CT) scans and cephalographs were taken at 10, 25, 42, and 84 days of age. Calvaria were harvested at 84 days of age for histomorphometric analysis. Mean differences were analyzed using a group by age analysis of variance. Analysis of the 3D CT scan data revealed that sites treated with TGF-β3 had significantly (P < .05) greater defect areas and significantly (P < .05) greater intracranial volumes through 84 days of age compared with controls. Histomorphometry showed that sites treated with TGF-β3 had patent suturectomy sites and significantly (P < .001) less new bone in the suturectomy site compared with controls. Serial radiograph data revealed significant (P < .05) differences in craniofacial growth from 25 to 84 days in TGF-β3-treated rabbits compared with controls. Data show that TGF-β3 administration delayed reossification and improved craniofacial growth in this rabbit model. These findings also suggest that this molecular-based therapy may have potential clinical use.

Tongue Morphology in Infant and Adult Bushbabies (Otolemur spp.)

Lingual fungiform papillae are the only structures on the anterior two-thirds of the tongue that contain taste receptor cells and are the first gustatory structures to encounter food items. In humans, density of fungiform papillae is associated with taste sensitivity and food selection. Nonhuman primates also use the sense of taste to detect the nutritional contents of potential food items. The present study examines the ontogeny, distribution, and density of fungiform papillae in two species of greater bushbaby that differ in dietary specialization. Using light and electron microscopic methods, adult and infant cadaveric specimens (n = 4) of the frugivorous Otolemur garnettii and the exudativorous O. crassicaudatus were examined. The density of fungiform papillae was measured in an additional three adults of each species using 0.5% methylene blue. Observations by light and scanning electron microscopy (SEM) indicate that receptor pores are open in neonates. One adult O. crassicaudatus examined by SEM shows a high number of anterior fungiform papillae on which one or more open taste pores can be observed. The mean density of fungiform papillae was more than 50% greater in O. garnettii (162.4 ± 70.63/cm2) compared to that in O. crassicaudatus (101.9 ± 20.63/cm2). These results suggest that greater bushbabies may have precocious ability to detect taste stimuli, based on the presence of open taste pores at birth. The apparent difference in fungiform papillae density between species requires further exploration.

How the Brain May Have Shaped Muscle Anatomy and Physiology: A Preliminary Study

Skeletal muscle fibers are often used to evaluate functional differences in locomotion. However, because there are energetic differences among muscle fiber cells, muscle fiber composition could be used to address evolutionary questions about energetics. Skeletal muscle is composed of two main types of fibers: Type I and II. The difference between the two can be reduced to how these muscle cells use oxygen and glucose. Type I fibers convert glucose to ATP using oxygen, while Type II fibers rely primarily on anaerobic metabolic processes. The expensive tissue hypothesis (ETH) proposes that the energetic demands imposed on the body by the brain result in a reduction in other expensive tissues (e.g., gastrointestinal tract). The original ETH dismisses the energetic demands of skeletal muscle, despite skeletal muscle being (1) an expensive tissue when active and (2) in direct competition for glucose with the brain. Based on these observations we hypothesize that larger brained primates will have relatively less muscle mass and a decrease in Type I fibers. As part of a larger study to test this hypothesis, we present data from 10 species of primates. We collected body mass, muscle mass, and biopsied four muscles from each specimen for histological procedures. We collected endocranial volumes from the literature. Using immunohistochemistry, a muscle fiber composition profile was created for each species sampled. Results show that larger brained primates have less muscle and fewer Type I fibers than primates with smaller brains. Results clarify the relationship between muscle mass and brain mass and illustrate how muscle mass could be used to address energetic questions. Anat Rec, 301:528–537, 2018.