Estimation of the forces exerted on the limb long bones of a white rhinoceros (Ceratotherium simum) using musculoskeletal modelling and simulation.

Heavy animals incur large forces on their limb bones, due to the transmission of body weight and ground reaction forces, and the contractions of the various muscles of the limbs. This is particularly true for rhinoceroses, the heaviest extant animals capable of galloping. Several studies have examined their musculoskeletal system and the forces their bones incur, but no detailed quantification has ever been attempted. Such quantification could help understand better the link between form and function in giant land animals. Here we constructed three-dimensional musculoskeletal models of the forelimb and hindlimb of Ceratotherium simum, the heaviest extant rhino species, and used static optimisation (inverse) simulations to estimate the forces applied on the bones when standing at rest, including magnitudes and directions. Overall, unsurprisingly, the most active muscles were antigravity muscles, which generate moments opposing body weight (thereby incurring the ground reaction force), and thus keep the joints extended, avoiding joint collapse via flexion. Some muscles have an antigravity action around several joints, and thus were found to be highly active, likely specialised in body weight support (ulnaris lateralis; digital flexors). The humerus was subjected to the greatest amount of forces in terms of total magnitude; forces on the humerus furthermore came from a great variety of directions. The radius was mainly subject to high-magnitude compressive joint reaction forces, but to little muscular tension, whereas the opposite pattern was observed for the ulna. The femur had a pattern similar to that of the humerus, and the tibia's pattern was intermediate, being subject to great compression in its caudal side but to great tension in its cranial side (i.e. bending). The fibula was subject to by far the lowest force magnitude. Overall, the forces estimated were consistent with the documented morphofunctional adaptations of C. simum's long bones, which have larger insertion areas for several muscles and a greater robusticity overall than those of lighter rhinos, likely reflecting the intense forces we estimated here. Our estimates of muscle and bone (joint) loading regimes for this giant tetrapod improve the understanding of the links between form and function in supportive tissues and could be extended to other aspects of bone morphology, such as microanatomy.

Regional variation of the cortical and trabecular bone material properties in the rabbit skull.

The material properties of some bones are known to vary with anatomical location, orientation and position within the bone (e.g., cortical and trabecular bone). Details of the heterogeneity and anisotropy of bone is an important consideration for biomechanical studies that apply techniques such as finite element analysis, as the outcomes will be influenced by the choice of material properties used. Datasets detailing the regional variation of material properties in the bones of the skull are sparse, leaving many finite element analyses of skulls no choice but to employ homogeneous, isotropic material properties, often using data from a different species to the one under investigation. Due to the growing significance of investigating the cranial biomechanics of the rabbit in basic science and clinical research, this study used nanoindentation to measure the elastic modulus of cortical and trabecular bone throughout the skull. The elastic moduli of cortical bone measured in the mediolateral and ventrodorsal direction were found to decrease posteriorly through the skull, while it was evenly distributed when measured in the anteroposterior direction. Furthermore, statistical tests showed that the variation of elastic moduli between separate regions (anterior, middle and posterior) of the skull were significantly different in cortical bone, but was not in trabecular bone. Elastic moduli measured in different orthotropic planes were also significantly different, with the moduli measured in the mediolateral direction consistently lower than that measured in either the anteroposterior or ventrodorsal direction. These findings demonstrate the significance of regional and directional variation in cortical bone elastic modulus, and therefore material properties in finite element models of the skull, particularly those of the rabbit, should consider the heterogeneous and orthotropic properties of skull bone when possible.

Airflow rates and breathlessness recovery from submaximal exercise in healthy adults: prospective, randomised, cross-over study.

OBJECTIVES: Facial airflow from a hand-held fan may reduce breathlessness severity and hasten postexertion recovery. Data from randomised controlled trials are limited and the optimal airflow speed remains unknown. We aimed to determine the effect of different airflow speeds on recovery from exercise-induced breathlessness. METHODS: A prospective, randomised, cross-over design. Ten healthy participants (seven male; mean age 29±4 years; height 175±9 cm; body mass 76.9±14.1 kg) completed six bouts of 4 min of exercise. During the first 5 min of a 20 min recovery phase, participants received one of five airflow speeds by holding a fan ~15 cm from their face, or no fan control, administered in random order. Fan A had an internal blade, and fan B had an external blade. Breathlessness was measured using a numerical rating scale (NRS) at minute intervals for the first 10 min, and facial skin temperature was recorded using a thermal imaging camera (immediately postexertion and 5 min recovery). RESULTS: Nine participants completed the trial. A significant main effect for airflow speed (p=0.016, ηp2=0.285) and interaction effect for airflow speed over time (p=0.008, ηp2=0.167) suggest that the airflow speed modifies breathlessness during recovery from exercise. Fan speeds of 1.7 m/s or greater increased the speed of recovery from breathlessness compared with control (p<0.05) with the highest airflow speeds (2.5 m/s and 3.3 m/s) giving greatest facial cooling. CONCLUSION: Higher airflow rates (1.7 m/s or greater) reduced self-reported recovery times from exercise-induced breathlessness and reduced facial temperature .

Functional reorganisation of the cranial skeleton during the cynodont-mammaliaform transition.

Skeletal simplification occurred in multiple vertebrate clades over the last 500 million years, including the evolution from premammalian cynodonts to mammals. This transition is characterised by the loss and reduction of cranial bones, the emergence of a novel jaw joint, and the rearrangement of the jaw musculature. These modifications have long been hypothesised to increase skull strength and efficiency during feeding. Here, we combine digital reconstruction and biomechanical modelling to show that there is no evidence for an increase in cranial strength and biomechanical performance. Our analyses demonstrate the selective functional reorganisation of the cranial skeleton, leading to reduced stresses in the braincase and the skull roof but increased stresses in the zygomatic region through this transition. This cranial functional reorganisation, reduction in mechanical advantage, and overall miniaturisation in body size are linked with a dietary specialisation to insectivory, permitting the subsequent morphological and ecological diversification of the mammalian lineage.

Assessment of the mechanical role of cranial sutures in the mammalian skull: Computational biomechanical modelling of the rat skull.

Cranial sutures are fibrocellular joints between the skull bones that are progressively replaced with bone throughout ontogeny, facilitating growth and cranial shape change. This transition from soft tissue to bone is reflected in the biomechanical properties of the craniofacial complex. However, the mechanical significance of cranial sutures has only been explored at a few localised areas within the mammalian skull, and as such our understanding of suture function in overall skull biomechanics is still limited. Here, we sought to determine how the overall strain environment is affected by the complex network of cranial sutures in the mammal skull. We combined two computational biomechanical methods, multibody dynamics analysis and finite element analysis, to simulate biting in a rat skull and compared models with and without cranial sutures. Our results show that including complex sutures in the rat model does not substantially change overall strain gradients across the cranium, particularly strain magnitudes in the bones overlying the brain. However, local variations in strain magnitudes and patterns can be observed in areas close to the sutures. These results show that, during feeding, sutures may be more important in some regions than others. Sutures should therefore be included in models that require accurate local strain magnitudes and patterns of cranial strain, particularly if models are developed for analysis of specific regions, such as the temporomandibular joint or zygomatic arch. Our results suggest that, for mammalian skulls, cranial sutures might be more important for allowing brain expansion during growth than redistributing biting loads across the cranium in adults.

Specific Sleep Health Domains as Predictors of Executive Function in Older Adults.

OBJECTIVES: Sleep health and executive function are multifaceted constructs that decline with age. Some evidence suggests that poor sleep health may underlie declines in executive function, but this relationship is not consistently found in cognitively normal older adults. The authors systematically investigated distinct sleep health domain associations with specific aspects of executive function. METHODS: Community-dwelling older adults completed clinical interviews, comprehensive neuropsychological assessments, and subjective sleep measures. Four sleep health domains were investigated: satisfaction/quality, sleep efficiency, sleep duration, and daytime sleepiness/fatigue. Hierarchical multiple regression analyses, adjusting for significant covariates, examined whether the sleep health domains differentially predicted executive function. RESULTS: Separate analyses found that greater sleep efficiency was associated with better response inhibition, while greater daytime sleepiness/fatigue was associated with worse cognitive flexibility. Categorical differences in sleep duration indicated that average durations, compared with short and long durations, had better executive function performance across measures. Sleep satisfaction/quality was not statistically associated with executive function. CONCLUSIONS: These findings have implications for sleep assessment and its intervention. Routine screening of sleep duration, efficiency, and daytime fatigue may be particularly useful in identifying those at greater risk of executive dysfunction. Targeting specific problems in sleep may serve to improve cognitive control and efficiency in older adults. Future research is warranted to establish the optimal hours of sleep duration for cognitive health.

Biomechanical evaluation of the unilateral crossbite on the asymmetrical development of the craniofacial complex. A mechano-morphological approach.

BACKGROUND AND OBJECTIVE: The occlusion effect on the craniofacial development is a controversial topic that has attracted the interest of many researchers but that remains unclear, mainly due to the difficulties on measure its mechanical response experimentally. This mechano-morphological relationship of the craniofacial growth is often explained by the periosteal and capsular matrices of the functional matrix hypothesis (FMH); however, its outcomes have not been analytically demonstrated yet. This computational study aims, therefore, to analytically demonstrate the mechano-morphological relationship in the craniofacial development of children with unilateral crossbite (UXB) using the finite element (FE) method. METHODS: The craniofacial complex asymmetry of ten children, five of whom exhibit UXB, was 3D-analysed and compared with the biomechanical response computed from a FE analysis of each patient's occlusion. Due to the complexity of the geometry and the multitude of contacts involved, the inherent limitations of the model were evaluated by comparing computed occlusal patterns with those recorded by an occlusal analysis on 3D printed copies. RESULTS: Comparison's outcomes proved the reliability of our models with just a deviation error below 6% between both approaches. Out of validation process, computational results showed that the significant elongation of mandibular branch in the contralateral side could be related to the mandibular shift and increase of thickness on the crossed side, and particularly of the posterior region. These morphological changes could be associated with periodontal overpressure (>4.7 kPa) and mandibular over deformation (0.002 ε) in that side, in agreement with the periosteal matrix's principles. Furthermore, the maxilla's transversal narrowing and the elevation of the maxillary and zygomatic regions on the crossed side were statistically demonstrated and seem to be related with their respective micro displacements at occlusion, as accounted by their specific capsule matrices. Our results were consistent with those reported clinically and demonstrated analytically the mechano-morphological relationship of children's craniofacial development based on the FMH's functional matrices. CONCLUSIONS: This study is a first step in the understanding of the occlusion's effect on the craniofacial development by computational methods. Our approach could help future engineers, researchers and clinicians to understand better the aetiology of some dental malocclusions and functional disorders improve the diagnosis or even predict the craniofacial development.

A hierarchical opportunistic screening model for osteoporosis using machine learning applied to clinical data and CT images.

BACKGROUND: Osteoporosis is a common metabolic skeletal disease and usually lacks obvious symptoms. Many individuals are not diagnosed until osteoporotic fractures occur. Bone mineral density (BMD) measured by dual-energy X-ray absorptiometry (DXA) is the gold standard for osteoporosis detection. However, only a limited percentage of people with osteoporosis risks undergo the DXA test. As a result, it is vital to develop methods to identify individuals at-risk based on methods other than DXA. RESULTS: We proposed a hierarchical model with three layers to detect osteoporosis using clinical data (including demographic characteristics and routine laboratory tests data) and CT images covering lumbar vertebral bodies rather than DXA data via machine learning. 2210 individuals over age 40 were collected retrospectively, among which 246 individuals' clinical data and CT images are both available. Irrelevant and redundant features were removed via statistical analysis. Consequently, 28 features, including 16 clinical data and 12 texture features demonstrated statistically significant differences (p < 0.05) between osteoporosis and normal groups. Six machine learning algorithms including logistic regression (LR), support vector machine with radial-basis function kernel, artificial neural network, random forests, eXtreme Gradient Boosting and Stacking that combined the above five classifiers were employed as classifiers to assess the performances of the model. Furthermore, to diminish the influence of data partitioning, the dataset was randomly split into training and test set with stratified sampling repeated five times. The results demonstrated that the hierarchical model based on LR showed better performances with an area under the receiver operating characteristic curve of 0.818, 0.838, and 0.962 for three layers, respectively in distinguishing individuals with osteoporosis and normal BMD. CONCLUSIONS: The proposed model showed great potential in opportunistic screening for osteoporosis without additional expense. It is hoped that this model could serve to detect osteoporosis as early as possible and thereby prevent serious complications of osteoporosis, such as osteoporosis fractures.

From micro to macroevolution: drivers of shape variation in an island radiation of Podarcis lizards.

Phenotypictraits have been shown to evolve in response to variation in the environment. However, the evolutionary processes underlying the emergence of phenotypic diversity can typically only be understood at the population level. Consequently, how subtle phenotypic differences at the intraspecific level can give rise to larger-scale changes in performance and ecology remains poorly understood. We here tested for the covariation between ecology, bite force, jaw muscle architecture, and the three-dimensional shape of the cranium and mandible in 16 insular populations of the lizards Podarcis melisellensis and P. sicula. We then compared the patterns observed at the among-population level with those observed at the interspecific level. We found that three-dimensional head shape as well as jaw musculature evolve similarly under similar ecological circumstances. Depending on the type of food consumed or on the level of sexual competition, different muscle groups were more developed and appeared to underlie changes in cranium and mandible shape. Our findings show that the local selective regimes are primary drivers of phenotypic variation resulting in predictable patterns of form and function. Moreover, intraspecific patterns of variation were generally consistent with those at the interspecific level, suggesting that microevolutionary variation may translate into macroevolutionary patterns of ecomorphological diversity.

Back to the bones: do muscle area assessment techniques predict functional evolution across a macroevolutionary radiation?

Measures of attachment or accommodation area on the skeleton are a popular means of rapidly generating estimates of muscle proportions and functional performance for use in large-scale macroevolutionary studies. Herein, we provide the first evaluation of the accuracy of these muscle area assessment (MAA) techniques for estimating muscle proportions, force outputs and bone loading in a comparative macroevolutionary context using the rodent masticatory system as a case study. We find that MAA approaches perform poorly, yielding large absolute errors in muscle properties, bite force and particularly bone stress. Perhaps more fundamentally, these methods regularly fail to correctly capture many qualitative differences between rodent morphotypes, particularly in stress patterns in finite-element models. Our findings cast doubts on the validity of these approaches as means to provide input data for biomechanical models applied to understand functional transitions in the fossil record, and perhaps even in taxon-rich statistical models that examine broad-scale macroevolutionary patterns. We suggest that future work should go back to the bones to test if correlations between attachment area and muscle size within homologous muscles across a large number of species yield strong predictive relationships that could be used to deliver more accurate predictions for macroevolutionary and functional studies.

Computational biomechanical modelling of the rabbit cranium during mastication.

Although a functional relationship between bone structure and mastication has been shown in some regions of the rabbit skull, the biomechanics of the whole cranium during mastication have yet to be fully explored. In terms of cranial biomechanics, the rabbit is a particularly interesting species due to its uniquely fenestrated rostrum, the mechanical function of which is debated. In addition, the rabbit processes food through incisor and molar biting within a single bite cycle, and the potential influence of these bite modes on skull biomechanics remains unknown. This study combined the in silico methods of multi-body dynamics and finite element analysis to compute musculoskeletal forces associated with a range of incisor and molar biting, and to predict the associated strains. The results show that the majority of the cranium, including the fenestrated rostrum, transmits masticatory strains. The peak strains generated over all bites were found to be attributed to both incisor and molar biting. This could be a consequence of a skull shape adapted to promote an even strain distribution for a combination of infrequent incisor bites and cyclic molar bites. However, some regions, such as the supraorbital process, experienced low peak strain for all masticatory loads considered, suggesting such regions are not designed to resist masticatory forces.

Regional Patterning in Tail Vertebral Form and Function in Chameleons (Chamaeleo calyptratus).

Previous studies have focused on documenting shape variation in the caudal vertebrae in chameleons underlying prehensile tail function. The goal of this study was to test the impact of this variation on tail function using multibody dynamic analysis (MDA). First, observations from dissections and 3D reconstructions generated from contrast-enhanced µCT scans were used to document regional variation in arrangement of the caudal muscles along the antero-posterior axis. Using MDA, we then tested the effect of vertebral shape geometry on biomechanical function. To address this question, four different MDA models were built: those with a distal vertebral shape and with either a distal or proximal musculature, and reciprocally the proximal vertebral shape with either the proximal or distal musculature. For each muscle configuration, we calculated the force required in each muscle group for the muscle force to balance an arbitrary external force applied to the model. The results showed that the models with a distal-type of musculature are the most efficient, regardless of vertebral shape. Our models also showed that the m. ilio-caudalis pars dorsalis is least efficient when combining the proximal vertebral shape and distal musculature, highlighting the importance of the length of the transverse process in combination with the lever-moment arm onto which muscle force is exerted. This initial model inevitably has a number of simplifications and assumptions, however its purpose is not to predict in vivo forces, but instead reveals the importance of vertebral shape and muscular arrangement on the total force the tail can generate, thus providing a better understanding of the biomechanical significance of the regional variations on tail grasping performance in chameleons.

Evolutionary biomechanics: hard tissues and soft evidence?

Biomechanical modelling is a powerful tool for quantifying the evolution of functional performance in extinct animals to understand key anatomical innovations and selective pressures driving major evolutionary radiations. However, the fossil record is composed predominantly of hard parts, forcing palaeontologists to reconstruct soft tissue properties in such models. Rarely are these reconstruction approaches validated on extant animals, despite soft tissue properties being highly determinant of functional performance. The extent to which soft tissue reconstructions and biomechanical models accurately predict quantitative or even qualitative patterns in macroevolutionary studies is therefore unknown. Here, we modelled the masticatory system in extant rodents to objectively test the ability of current muscle reconstruction methods to correctly identify quantitative and qualitative differences between macroevolutionary morphotypes. Baseline models generated using measured soft tissue properties yielded differences in muscle proportions, bite force, and bone stress expected between extant sciuromorph, myomorph, and hystricomorph rodents. However, predictions from models generated using reconstruction methods typically used in fossil studies varied widely from high levels of quantitative accuracy to a failure to correctly capture even relative differences between macroevolutionary morphotypes. Our novel experiment emphasizes that correctly reconstructing even qualitative differences between taxa in a macroevolutionary radiation is challenging using current methods. Future studies of fossil taxa should incorporate systematic assessments of reconstruction error into their hypothesis testing and, moreover, seek to expand primary datasets on muscle properties in extant taxa to better inform soft tissue reconstructions in macroevolutionary studies.

Comparative cranial biomechanics in two lizard species: impact of variation in cranial design.

Cranial morphology in lepidosaurs is highly disparate and characterised by the frequent loss or reduction of bony elements. In varanids and geckos, the loss of the postorbital bar is associated with changes in skull shape, but the mechanical principles underlying this variation remain poorly understood. Here, we sought to determine how the overall cranial architecture and the presence of the postorbital bar relate to the loading and deformation of the cranial bones during biting in lepidosaurs. Using computer-based simulation techniques, we compared cranial biomechanics in the varanid Varanus niloticus and the teiid Salvator merianae, two large, active foragers. The overall strain magnitude and distribution across the cranium were similar in the two species, despite lower strain gradients in V. niloticus In S. merianae, the postorbital bar is important for resistance of the cranium to feeding loads. The postorbital ligament, which in varanids partially replaces the postorbital bar, does not affect bone strain. Our results suggest that the reduction of the postorbital bar impaired neither biting performance nor the structural resistance of the cranium to feeding loads in V. niloticus Differences in bone strain between the two species might reflect demands imposed by feeding and non-feeding functions on cranial shape. Beyond variation in cranial bone strain related to species-specific morphological differences, our results reveal that similar mechanical behaviour is shared by lizards with distinct cranial shapes. Contrary to the situation in mammals, the morphology of the circumorbital region, calvaria and palate appears to be important for withstanding high feeding loads in these lizards.

Mathematical modeling of canonical and non-canonical NF-κB pathways in TNF stimulation.

BACKGROUND AND OBJECTIVE: NF-κB can be activated by the canonical and non-canonical pathways. These two pathways interplay via the TRAF1|NIK complex after stimulation by TNF. However existing mathematical models of two pathways are inadequate. In this context, an improved mathematical model is constructed to simulate these two pathways and their coupling stimulated by TNF. METHODS: A schematic description of two NF-κB pathways and their relation after TNF stimulation is constructed at first. Then twenty-eight ordinary differential equations are utilized to build the mathematical model. Model equations are solved via the ordinary differential equation solver (ode23). RESULTS: The proposed model firstly reconstructs the changes in concentrations of NF-κB pathway related biochemical factors with time, and further investigates the underlying mechanism of interaction between two pathways through the TRAF1|NIK complex after stimulation. CONCLUSIONS: The model is validated through good agreement between simulation results and published experimental observations. This study helps to well understand the canonical and non-canonical pathways and their interaction. It also provides a potential tool to investigate how the dysregulated pathways act in pathological conditions.

The influence of musculoskeletal forces on the growth of the prenatal cortex in the ilium: a finite element study.

Remodelling and adaptation of bone within the pelvis is believed to be influenced by the mechanical strains generated during locomotion. Variation in the cortical bone thickness observed in the prenatal ilium has been linked to the musculoskeletal loading associated with in utero movements; for example the development of a thicker gluteal cortex is a possible response to contractions of the gluteal muscles. This study examines if the strains generated in the prenatal iliac cortex due to musculoskeletal loading in utero are capable of initiating bone remodelling to either maintain homeostasis or form new bone. Computational modelling techniques were used firstly to predict the muscle forces and resultant joint reaction force acting on the pelvis during a range of in utero movements. Finite element analyses were subsequently performed to calculate the von Mises strains induced in the prenatal ilium. The results demonstrated that strains generated in the iliac cortex were above the thresholds suggested to regulate bone remodelling to either maintain homeostasis or form new bone. Further simulations are required to investigate the extent to which the heterogeneous cortex forms in response to these strains (i.e., remodelling) or if developmental bone modelling plays a more pivotal role.

Towards an early 3D-diagnosis of craniofacial asymmetry by computing the accurate midplane: A PCA-based method.

BACKGROUND AND OBJECTIVE: Craniofacial asymmetry is a common growth disorder often caused by unilateral chewing. Although an early orthodontic treatment would avoid surgical procedures later in life, the uncertainty of defining the accurate sagittal midplane potentially leads to misdiagnosis and therefore inaccurate orthodontic treatment plans. This novel study aims to 3D-diagnose craniofacial complex malformations in children with unilateral crossbite (UXB) considering a midplane which compensates the asymmetric morphology. METHODS: The sagittal midplane of 20 children, fifteen of whom exhibited UXB, was computed by a PCA-based method which compensates the asymmetry mirroring the 3D models obtained from cone-beam computed tomography data. Once determined, one side of the data was mirrored using the computed midplane to visualize the malformations on the hard and soft tissues by 3D-computing the distances between both halves. Additionally, 31 skull's landmarks were manually placed in each model to study the principal variation modes and the significant differences in the group of subjects with and without UXB through PCA and Mann-Whitney U test analyses respectively. RESULTS: Morphological 3D-analysis showed pronounced deformities and aesthetic implications for patients with severe asymmetry (jaw deviation > 0.8 mm) in whole craniofacial system, while initial signs of asymmetry were found indistinctly in the mandible or maxilla. We detected significant (p < 0.05) malformations for example in mandibular ramus length (0.0086), maxillary palate width (0.0481) and condylar head width (0.0408). Craniofacial malformations increased the landmarks' variability in the group of patients with UXB over the control group requiring 8 variation modes more to define 99% of the sample' variability. CONCLUSIONS: Our findings demonstrated the viability of early diagnosis of craniofacial asymmetry through computing the accurate sagittal midplane which compensates the individual's asymmetrical morphology. Furthermore, this study provides important computational insights into the determination of craniofacial deformities which are caused by UXB, following some empirical findings of previous clinical studies. Hence, this computational approach can be useful for the development of new software in craniofacial surgery or for its use in biomedical research and clinical practice.

A 3D cephalometric protocol for the accurate quantification of the craniofacial symmetry and facial growth.

BACKGROUND: Cephalometric analysis is used to evaluate facial growth, to study the anatomical relationships within the face. Cephalometric assessment is based on 2D radiographic images, either the sagittal or coronal planes and is an inherently inaccurate methodology. The wide availability of 3D imaging techniques, such as computed tomography and magnetic resonance imaging make routine 3D analysis of facial morphology feasible. 3D cephalometry may not only provide a more accurate quantification of the craniofacial morphology and longitudinal growth, but also the differentiation of subtle changes in occlusion. However, a reliable protocol for the computation of craniofacial symmetry and quantification of craniofacial morphology is still a topic of extensive research. Here, a protocol for 3D cephalometric analysis for both the identification of the natural head position (NHP) and the accurate quantification of facial growth and facial asymmetry is proposed and evaluated. A phantom study was conducted to assess the performance of the protocol and to quantify the ability to repeatedly and reliably align skulls with the NHP and quantify the degree of accuracy with which facial growth and facial asymmetry can be measured. RESULTS: The results obtained show that the protocol allows consistent alignment with the NHP, with an overall average error (and standard deviation) of just 0.17 (9.10e-6) mm, with variations of 0.21 (2.77e-17) mm in the frontonasal suture and 0.30 (5.55e-17) mm in the most prominent point in the chin. The average errors associated with simulated facial growth ranged from 1.83 to 3.75% for 2 years' growth and from - 9.57 to 14.69% for 4 years, while the error in the quantification of facial asymmetry ranged from - 11.38 to 9.31%. CONCLUSIONS: The protocol for 3D skull alignment produces accurate and landmark free estimation of the true symmetry of the head. It allows a reliable alignment of the skull in the NHP independently of user-defined landmarks, as well as an accurate quantification of facial growth and asymmetry.

Neurocranial development of the coelacanth and the evolution of the sarcopterygian head.

The neurocranium of sarcopterygian fishes was originally divided into an anterior (ethmosphenoid) and posterior (otoccipital) portion by an intracranial joint, and underwent major changes in its overall geometry before fusing into a single unit in lungfishes and early tetrapods1. Although the pattern of these changes is well-documented, the developmental mechanisms that underpin variation in the form of the neurocranium and its associated soft tissues during the evolution of sarcopterygian fishes remain poorly understood. The coelacanth Latimeria is the only known living vertebrate that retains an intracranial joint2,3. Despite its importance for understanding neurocranial evolution, the development of the neurocranium of this ovoviviparous fish remains unknown. Here we investigate the ontogeny of the neurocranium and brain in Latimeria chalumnae using conventional and synchrotron X-ray micro-computed tomography as well as magnetic resonance imaging, performed on an extensive growth series for this species. We describe the neurocranium at the earliest developmental stage known for Latimeria, as well as the major changes that the neurocranium undergoes during ontogeny. Changes in the neurocranium are associated with an extreme reduction in the relative size of the brain along with an enlargement of the notochord. The development of the notochord appears to have a major effect on the surrounding cranial components, and might underpin the formation of the intracranial joint. Our results shed light on the interplay between the neurocranium and its adjacent soft tissues during development in Latimeria, and provide insights into the developmental mechanisms that are likely to have underpinned the evolution of neurocranial diversity in sarcopterygian fishes.