A skew in poo: Biases in primate fecal isotope analysis and recommendations for standardized sample preparation.

Feces are a treasure trove in the study of animal behavior and ecology. Stable carbon and nitrogen isotope analysis allows to assess the dietary niches of elusive primate species and primate breastfeeding behavior. However, some fecal isotope data may unwillingly be biased toward the isotope ratios of undigested plant matter, requiring more consistent sample preparation protocols. We assess the impact of this potential data skew in 114 fecal samples of wild bonobos (Pan paniscus) by measuring the isotope differences (Δ13 C, Δ15 N) between bulk fecal samples containing larger particles (>1 mm) and filtered samples containing only small particles (<1 mm). We assess the influence of fecal carbon and nitrogen content (ΔC:N) and sample donor age (subadult, adult) on the resulting Δ13 C, Δ15 N values (n = 228). Additionally, we measure the isotope ratios in three systematically sieved fecal samples of chimpanzees (Pan troglodytes verus), with particle sizes ranging from 20 µm to 8 mm (n = 30). We found differences in fecal carbon and nitrogen content, with the smaller fecal fraction containing more nitrogen on average. While the Δ13 C values were small and not affected by age or ΔC:N, the Δ15 N values were significantly influenced by fecal ΔC:N, possibly resulting from the differing proportions of undigested plant macroparticles. Significant relationships between carbon stable isotope ratios (δ13 C) values and %C in large fecal fractions of both age groups corroborated this assessment. Δ15 N values were significantly larger in adults than subadults, which should be of concern in isotope studies comparing adult females with infants to assess breastfeeding. We found a random variation of up to 3.0‰ in δ13 C and 2.0‰ in nitrogen stable isotope ratios within the chimpanzee fecal samples separated by particle sizes. We show that particle size influences isotope ratios and propose a simple, cost-effective filtration method for primate feces to exclude larger undigested food particles from the analysis, which can easily be adopted by labs worldwide.

Unilateral Hypofunction of the Masseter Leads to Molecular and 3D Morphometric Signs of Atrophy in Ipsilateral Agonist Masticatory Muscles in Adult Mice.

Mice are commonly used to study mandibular dynamics due to their similarity in chewing cycle patterns with humans. Adult mice treated unilaterally with botulinum toxin type A (BoNTA) in the masseter exhibit atrophy of this muscle characterized by an increase in the gene expression of atrophy-related molecular markers, and a reduction in both muscle fiber diameter and muscle mass at 14d. However, the impact of this muscle imbalance on the non-treated masticatory muscles remains unexplored. Here, we hypothesize that the unilateral masseter hypofunction leads to molecular and 3D morphometric signs of atrophy of the masseter and its agonist masticatory muscles in adult mice. Twenty-three 8-week-old male BALB/c mice received a single injection of BoNTA in the right masseter, whereas the left masseter received the same volume of saline solution (control side). Animals were euthanized at 2d, 7d, and 14d, and the masticatory muscles were analyzed for mRNA expression. Five heads were harvested at 14d, fixed, stained with a contrast-enhanced agent, and scanned using X-ray microtomography. The three-dimensional morphometric parameters (the volume and thickness) from muscles in situ were obtained. Atrogin-1/MAFbx, MuRF-1, and Myogenin mRNA gene expression were significantly increased at 2 and 7d for both the masseter and temporalis from the BoNTA side. For medial pterygoid, increased mRNA gene expression was found at 7d for Atrogin-1/MAFbx and at 2d-7d for Myogenin. Both the volume and thickness of the masseter, temporalis, and medial pterygoid muscles from the BoNTA side were significantly reduced at 14d. In contrast, the lateral pterygoid from the BoNTA side showed a significant increase in volume at 14d. Therefore, the unilateral hypofunction of the masseter leads to molecular and morphological signs of atrophy in both the BoNTA-injected muscle and its agonistic non-injected masticatory muscles. The generalized effect on the mouse masticatory apparatus when one of its components is intervened suggests the need for more clinical studies to determine the safety of BoNTA usage in clinical dentistry.

Movement analysis of primate molar teeth under load using synchrotron X-ray microtomography.

Mammalian teeth have to sustain repetitive and high chewing loads without failure. Key to this capability is the periodontal ligament (PDL), a connective tissue containing a collagenous fibre network which connects the tooth roots to the alveolar bone socket and which allows the teeth to move when loaded. It has been suggested that rodent molars under load experience a screw-like downward motion but it remains unclear whether this movement also occurs in primates. Here we use synchroton micro-computed tomography paired with an axial loading setup to investigate the form-function relationship between tooth movement and the morphology of the PDL space in a non-human primate, the mouse lemur (Microcebus murinus). The loading behavior of both mandibular and maxillary molars showed a three-dimensional movement with translational and rotational components, which pushes the tooth into the alveolar socket. Moreover, we found a non-uniform PDL thickness distribution and a gradual increase in volumetric proportion of the periodontal vasculature from cervical to apical. Our results suggest that the PDL morphology may optimize the three-dimensional tooth movement to avoid high stresses under loading.

Quantifying maxillary development in chimpanzees and humans: An analysis of prognathism and orthognathism at the morphological and microscopic scales.

Facial orientation (projection and degree of prognathism) and form in hominins is highly variable, likely related to evolutionary modifications of the microscopic process of bone modeling (the simultaneous cellular activities of bone formation and resorption) during ontogeny. However, in anteriorly projected faces such as those of early hominins, little is known about the link between bone modeling and facial developmental patterns. Similarly, these aspects have been infrequently investigated in extant great apes. In this study, quantitative methods were applied to a cross-sectional ontogenetic sample of 33 chimpanzees (Pan troglodytes verus) and 59 modern humans (Homo sapiens) to compare the development of maxillary prognathism to orthognathism at both microscopic and macroscopic (or morphological) scales using surface histology and geometric morphometric techniques. Chimpanzees express on average lower amounts of bone resorption than humans on the maxillary periosteum throughout ontogeny; however, the premaxilla is consistently resorbed from early stages on. The presence of bone resorption in the chimpanzee premaxilla, such as that seen in some early hominins, suggests a more ape-like pattern of maxillary bone modeling in these specimens. However, this shows that similarities in bone modeling patterns can lead to variations in shape, suggesting that other aspects of facial growth (such as modifications of rates and timings of development, as well as sutural growth) also played a crucial role in facial evolution.

The effect of high wear diets on the relative pulp volume of the lower molars.

OBJECTIVES: One role of dental pulp is in the upkeep and maintenance of dentine. Under wear, odontoblasts in the pulp deposit tertiary dentine to ensure the sensitive internal dental tissues are not exposed and vulnerable to infection. It follows that there may be an adaptive advantage for increasing molar pulp volume in anthropoid primate taxa that are prone to high levels of wear. The relative volume of dental pulp is therefore predicted to covary with dietary abrasiveness (in the sense of including foods that cause high degrees of wear). MATERIALS AND METHODS: We examined relatively unworn lower second molars in pairs of species of extant hominoids, cebids, and pitheciids that vary in the abrasiveness of their diet (n = 36). Using micro-CT scans, we measured the percent of tooth that is pulp (PTP) as the ratio of pulp volume to that of the total volume of the tooth. RESULTS: We found that in each pair of species, the taxa that consume a more abrasive diet had a significantly higher PTP than the closely related taxa that consume a softer diet. CONCLUSIONS: Our results point to an adaptive mechanism in the molars of taxa that consume abrasive diets and are thus subject to higher levels of wear. Our results provide additional understanding of the relationship between dental pulp and diet and may offer insight into the diet of extinct taxa such as Paranthropus boisei or into the adaptive context of the taurodont molars of Neanderthals.

Elevated activity levels do not influence extrinsic fiber attachment morphology on the surface of muscle-attachment sites.

Extrinsic fibers (EFs) are a type of penetrating collagenous fiber, closely related to the periodontal ligament, which help anchor soft tissue into bone. These fibers are associated with muscle attachment sites (entheses). Their size and grouping patterns are thought to be indicative of the loading history of the muscle. EFs are of particular significance in anthropology as potential tools for the reconstruction of behavior from skeletal remains and, specifically, entheses. In this study, we used a mouse model to experimentally test how activity level alters the morphology of EF insertion sites on the bone surface of a fibrocartilaginous enthesis, the biceps brachii insertion. Further, we adapted surface metrological techniques from studies of dental wear to perform automated, quantitative and non-destructive analysis of bone surface histology. Our results show that experimentally increased activity had no significant effect on the quantity or density of EF insertions at the enthesis, nor on the size of those insertions. Although EF presence does indicate muscle attachment, activity did not have an observable effect on EF morphology.

The Middle Pleistocene hominin mandible from Payre (Ardèche, France).

Although Neandertals are the best-known fossil hominins, the tempo and evolutionary processes in their lineage are strongly debated. This is in part due to the scarcity of the fossil record, in particular before the marine isotopic stage (MIS) 5. In 2010, a partial hominin mandible was discovered at the Middle Paleolithic site of Payre (France) in a layer that is dated to the end of MIS 8/beginning of MIS 7, a time period for which very few fossils are known in Europe. The Payre 15 mandible retains the complete symphyseal region and right lateral corpus with heavily worn P4, M1, and M2 in situ. Taphonomic modifications in the form of three notches suggest that this individual was chewed by a carnivore. We provide here the first detailed description of this specimen and a comparative analysis that includes morphological features, linear mandibular dimensions, an elliptic Fourier analysis of the symphysis, and a morphometric analysis of the M1 roots (based on segmented CT scan data). Our comparative sample encompasses European Middle and Upper Pleistocene specimens attributed to Homo heidelbergensis and Homo neanderthalensis, Upper Pleistocene Homo sapiens, and Holocene Homo sapiens. The Payre 15 mandible shows a combination of primitive and Neandertal-like features, with a receding symphyseal profile without any element of the mentum osseum, a posterior location of the mental foramen and lateral prominence. Its mandibular body is tall and thick anteriorly. Payre 15 has mesotaurodont M1 roots and a three-rooted M2. By its dimensions and combination of features, Payre 15 aligns better with Middle Pleistocene European hominins than with MIS 6-3 Neandertals. Noteworthy, it falls well within the range of variation of the Sima de los Huesos sample. Our results underscore that the total pattern of Neandertal-derived morphology was not achieved at the beginning of the MIS 7 and suggest a low level of mandibular diachronic changes for the period MIS 11-7.

Intraspecific variability in human maxillary bone modeling patterns during ontogeny.

OBJECTIVES: This study compares the ontogenetic bone modeling patterns of the maxilla to the related morphological changes in three human populations to better understand how morphological variability within a species is established during ontogeny at both micro- and macroscopic levels. MATERIALS AND METHODS: The maxillary bones of an ontogenetic sample of 145 subadult and adult individuals from Greenland (Inuit), Western Europe (France, Germany, and Portugal), and South Africa (Khoekhoe and San) were analyzed. Bone formation and resorption were quantified using histological methods to visualize the bone modeling patterns. In parallel, semilandmark geometric morphometric techniques were used on 3D models of the same individuals to capture the morphological changes. Multivariate statistics were applied and shape differences between age groups were visualized through heat maps. RESULTS: The three populations show differences in the degree of shape change acquired during ontogeny, leading to divergences in the developmental trajectories. Only subtle population differences in the bone modeling patterns were found, which were maintained throughout ontogeny. Bone resorption in adults mirrors the pattern found in subadults, but is expressed at lower intensities. DISCUSSION: Our data demonstrate that maxillary morphological differences observed in three geographically distinct human populations are also reflected at the microscopic scale. However, we suggest that these differences are mostly driven by changes in rates and timings of the cellular activities, as only slight discrepancies in the location of bone resorption could be observed. The shared general bone modeling pattern is likely characteristic of all Homo sapiens, and can be observed throughout ontogeny.

Mineral Deposits in Ficus Leaves: Morphologies and Locations in Relation to Function.

Ficus trees are adapted to diverse environments and have some of the highest rates of photosynthesis among trees. Ficus leaves can deposit one or more of the three major mineral types found in leaves: amorphous calcium carbonate cystoliths, calcium oxalates, and silica phytoliths. In order to better understand the functions of these minerals and the control that the leaf exerts over mineral deposition, we investigated leaves from 10 Ficus species from vastly different environments (Rehovot, Israel; Bologna, Italy; Issa Valley, Tanzania; and Ngogo, Uganda). We identified the mineral locations in the soft tissues, the relative distributions of the minerals, and mineral volume contents using microcomputed tomography. Each Ficus species is characterized by a unique 3D mineral distribution that is preserved in different environments. The mineral distribution patterns are generally different on the adaxial and abaxial sides of the leaf. All species examined have abundant calcium oxalate deposits around the veins. We used micromodulated fluorimetry to examine the effect of cystoliths on photosynthetic efficiency in two species having cystoliths abaxially and adaxially (Ficusmicrocarpa) or only abaxially (Ficuscarica). In F. microcarpa, both adaxial and abaxial cystoliths efficiently contributed to light redistribution inside the leaf and, hence, increased photosynthetic efficiency, whereas in F. carica, the abaxial cystoliths did not increase photosynthetic efficiency.

Ontogeny of the human maxilla: a study of intra-population variability combining surface bone histology and geometric morphometrics.

Bone modeling is the process by which bone grows in size and models its shape via the cellular activities of the osteoblasts and osteoclasts that respectively form and remove bone. The patterns of expression of these two activities, visible on bone surfaces, are poorly understood during facial ontogeny in Homo sapiens; this is due mainly to small sample sizes and a lack of quantitative data. Furthermore, how microscopic activities are related to the development of morphological features, like the uniquely human-canine fossa, has been rarely explored. We developed novel techniques for quantifying and visualizing variability in bone modeling patterns and applied these methods to the human maxilla to better understand its development at the micro- and macroscopic levels. We used a cross-sectional ontogenetic series of 47 skulls of known calendar age, ranging from birth to 12 years, from a population of European ancestry. Surface histology was employed to record and quantify formation and resorption on the maxilla, and digital maps representing each individual's bone modeling patterns were created. Semilandmark geometric morphometric (GM) methods and multivariate statistics were used to analyze facial growth. Our results demonstrate that surface histology and GM methods give complementary results, and can be used as an integrative approach in ontogenetic studies. The bone modeling patterns specific to our sample are expressed early in ontogeny, and fairly constant through time. Bone resorption varies in the size of its fields, but not in location. Consequently, absence of bone resorption in extinct species with small sample sizes should be interpreted with caution. At the macroscopic level, maxillary growth is predominant in the top half of the bone where bone formation is mostly present. Our results suggest that maxillary growth in humans is highly constrained from early stages in ontogeny, and morphological changes are likely driven by changes in osteoblastic and osteoclastic rates of expression rather than differences in the bone modeling patterns (i.e. changes in location of formation and resorption). Finally, the results of the micro- and macroscopic analyses suggest that the development of the canine fossa results from a combination of bone resorption and bone growth in the surrounding region.

Mandibular molar root and pulp cavity morphology in Homo naledi and other Plio-Pleistocene hominins.

The craniomandibular morphology of Homo naledi shows variable resemblances with species across Homo, which confounds an easy assessment of its phylogenetic position. In terms of skull shape, H. naledi has its closest affinities with Homo erectus, while mandibular shape places it closer to early Homo. From a tooth crown perspective, the smaller molars of H. naledi make it distinct from early Homo and H. erectus. Here, we compare the mandibular molar root morphology of six H. naledi individuals from the Dinaledi Chamber to those of African and Eurasian Plio-Pleistocene fossil hominins (totalling 183 mandibular first, second and third molars). The analysis of five root metric variables (cervical plane area, root length, root cervix volume, root branch volume, and root surface area) derived from microCT reconstructions reveals that the molar roots of H. naledi are smaller than those of Homo habilis, Homo rudolfensis, and H. erectus, but that they resemble those of three Homo sp. specimens from Swartkrans and Koobi Fora in size and overall appearance. Moreover, though H. naledi molar roots are similar in absolute size to Pleistocene Homo sapiens, they differ from H. sapiens in having a larger root volume for a given cervical plane area and less taurodont roots; the root cervix-to-branch proportions of H. naledi are comparable to those of Australopithecus africanus and species of Paranthropus. H. naledi also shares a metameric root volume pattern (M2 > M3 > M1) with Australopithecus and Paranthropus but not with any of the other Homo species (M2 > M1 > M3). Our findings therefore concur with previous studies that found that H. naledi shares plesiomorphic features with early Homo, Australopithecus, and Paranthropus. While absolute molar root size aligns H. naledi with Homo from North and South Africa, it is distinguishable from these in terms of root volumetric proportions.

Unexpected hard-object feeding in Western lowland gorillas.

OBJECTIVES: Gorilla diets are characterized by large amounts of fruit and tough fibrous plant material. Hard-object feeding is not generally associated with this genus as the high crests on their molar teeth would be at risk of damage from the mechanically challenging woody endocarp. This study aims to demonstrate that at least one population of western lowland gorillas are seasonal hard-object feeders, orally processing the seeds of Coula edulis. MATERIALS AND METHODS: Feeding behavior of habituated western lowland gorillas and phenology of fruiting trees was observed over a 4-year period to determine the extent they exploited the seeds of C. edulis. Additionally, the endocarps of C. edulis were subjected to testing to determine their mechanical properties. RESULTS: Our results demonstrate that during the fruiting season (January, February, and December) gorillas consistently opened the seeds of C. edulis using their postcanine dentition. The protective endocarp is composed of a very stiff material, presenting a substantial mechanical challenge to a gorilla. However, the high ratio between elastic modulus and toughness will facilitate brittle, cataclysmic fracture of the seed shell given a high enough load. DISCUSSION: Although a rich energy source, C. edulis likely tax gorilla dentitions to their upper limit. The rarity of such behavior at sites where it could be observed may indicate a degree of social learning or culture driving its occurrence. This shows a greater breadth of gorilla diets than previously described and suggests gorillas may be a useful model for interpreting the dietary mechanics that necessitated robust craniodental morphology in australopiths.

Dust affects chewing efficiency and tooth wear in forest dwelling Western chimpanzees (Pan troglodytes verus).

OBJECTIVES: In humans it has been shown that abrasive particles in the diet result in increased tooth wear and less intense chewing behavior, both of which decrease chewing efficiency. This behavioral response may also exist in non-human primates as a means to reduce the wear effect of dust-laden food. Here we tested whether the periodical occurrence of abrasive dust particles in the diet of Western chimpanzees affects tooth wear and reduces chewing efficiency. MATERIALS AND METHODS: We measured fecal particle size of undigested food matter as an indicator of chewing efficiency in 13 Western chimpanzees of the Taï National Park (Ivory Coast) before (wet), after (wet) and during a dust-rich (dry) period. Moreover, feeding data were compiled for a further 12 chimpanzees and matched to three-dimensional surface texture data measured on two molar facets of 26 skulls of the same population. RESULTS: Fecal particles were larger during the dry period, indicating a reduced chewing efficiency compared to wet periods; age and sex did not have an effect. Concomitantly, dust led to an increase of abrasive wear evidenced by smaller texture features and higher density of fine furrows on wear facets. DISCUSSION: Our findings show that a periodical increase in dust loads on foods places a dietary-physiological stress on the digestive system in chimpanzees. We suggest that the impact of extrinsic abrasive particles from globally acting periodical dust-laden winds may affect evolutionary fitness. Further studies are required to elucidate this relationship in other non-human primates and fossil hominins.

Masseter muscle atrophy impairs bone quality of the mandibular condyle but not the alveolar process early after induction.

BACKGROUND: Masseter muscle function influences mandibular bone homeostasis. As previously reported, bone resorption markers increased in the mouse mandibular condyle two days after masseter paralysis induced with botulinum toxin type A (BoNTA), followed by local bone loss. OBJECTIVE: This study aimed to evaluate the bone quality of both the mandibular condyle and alveolar process in the mandible of adult mice during the early stage of a BoNTA-induced masseter muscle atrophy, using a combined 3D histomorphometrics and shape analysis approach. METHODS: Adult BALB/c mice were divided into an untreated control group and an experimental group; the latter received one single BoNTA injection in the right masseter (BoNTA-right) and saline in the left masseter (Saline-left). 3D bone microstructural changes in the mandibular condyle and alveolar process were determined with high-resolution microtomography. Additionally, landmark-based geometric morphometrics was implemented to assess external shape changes. RESULTS: After 2 weeks, masseter mass was significantly reduced (P-value <0.001). When compared to Saline-left and untreated condyles, BoNTA-right condyles showed significant bone loss (P-value <0.001) and shape changes. No significant bone loss was observed in the alveolar processes of any of the groups (P-value >0.05). CONCLUSION: Condyle bone quality deteriorates at an early stage of BoNTA-induced masseter muscle atrophy, and before the alveolar process is affected. Since the observed bone microstructural changes resemble those in human temporomandibular joint degenerative disorders, the clinical safety of BoNTA intervention in the masticatory apparatus remains to be clarified.

Dental topography and the diet of Homo naledi.

Though late Middle Pleistocene in age, Homo naledi is characterized by a mosaic of Australopithecus-like (e.g., curved fingers, small brains) and Homo-like (e.g., elongated lower limbs) traits, which may suggest it occupied a unique ecological niche. Ecological reconstructions inform on niche occupation, and are particularly successful when using dental material. Tooth shape (via dental topography) and size were quantified for four groups of South African Plio-Pleistocene hominins (specimens of Australopithecus africanus, Paranthropus robustus, H. naledi, and Homo sp.) on relatively unworn M2s to investigate possible ecological differentiation in H. naledi relative to taxa with similar known geographical ranges. H. naledi has smaller, but higher-crowned and more wear resistant teeth than Australopithecus and Paranthropus. These results are found in both lightly and moderately worn teeth. There are no differences in tooth sharpness or complexity. Combined with the high level of dental chipping in H. naledi, this suggests that, relative to Australopithecus and Paranthropus, H. naledi consumed foods with similar fracture mechanics properties but more abrasive particles (e.g., dust, grit), which could be due to a dietary and/or environmental shift(s). The same factors that differentiate H. naledi from Australopithecus and Paranthropus may also differentiate it from Homo sp., which geologically predates it, in the same way. Compared to the great apes, all hominins have sharper teeth, indicating they consumed foods requiring higher shear forces during mastication. Despite some anatomical similarities, H. naledi likely occupied a distinct ecological niche from the South African hominins that predate it.

Ontogenetic changes to muscle architectural properties within the jaw-adductor musculature of Macaca fascicularis.

OBJECTIVES: Changes to soft- and hard-tissue components of the masticatory complex during development can impact functional performance by altering muscle excursion potential, maximum muscle forces, and the efficiency of force transfer to specific bitepoints. Within Macaca fascicularis, older individuals exploit larger, more mechanically resistant food items and more frequently utilize wide-gape jaw postures. We therefore predict that key architectural and biomechanical variables will scale during ontogeny to maximize bite force and gape potential within older, larger-bodied individuals. MATERIALS AND METHODS: We analyzed 26 specimens of M. fascicularis, representing a full developmental spectrum. The temporalis, superficial masseter, and deep masseter were dissected to determine muscle mass, fiber length, and physiologic cross-sectional area (PCSA). Lever-arm lengths were also measured for each muscle, alongside the height of the temporomandibular joint (TMJ) and basicranial length. These variables were scaled against two biomechanical variables (jaw length and condyle-molar length) to determine relative developmental changes within these parameters. RESULTS: During ontogeny, muscle mass, fiber length, and PCSA scaled with positive allometry relative to jaw length and condyle-molar length within all muscles. TMJ height also scaled with positive allometry, while muscle lever arms scaled with isometry relative to jaw length and with positive allometry (temporalis) or isometry (superficial and deep masseter) relative to condyle-molar length. CONCLUSION: Larger individuals demonstrate adaptations during development towards maximizing gape potential and bite force potential at both an anterior and posterior bitepoint. These data provide anatomical evidence to support field observations of dietary and behavioral differences between juvenile and adult M. fascicularis.

Congenital muscle dystrophy and diet consistency affect mouse skull shape differently.

The bones of the mammalian skull respond plastically to changes in masticatory function. However, the extent to which muscle function affects the growth and development of the skull, whose regions have different maturity patterns, remains unclear. Using muscle dissection and 3D landmark-based geometric morphometrics we investigated the effect of changes in muscle function established either before or after weaning, on skull shape and muscle mass in adult mice. We compared temporalis and masseter mass and skull shape in mice with a congenital muscle dystrophy (mdx) and wild type (wt) mice fed on either a hard or a soft diet. We found that dystrophy and diet have distinct effects on the morphology of the skull and the masticatory muscles. Mdx mice show a flattened neurocranium with a more dorsally displaced foramen magnum and an anteriorly placed mandibular condyle compared with wt mice. Compared with hard diet mice, soft diet mice had lower masseter mass and a face with more gracile features as well as labially inclined incisors, suggesting reduced bite strength. Thus, while the early-maturing neurocranium and the posterior portion of the mandible are affected by the congenital dystrophy, the late-maturing face including the anterior part of the mandible responds to dietary differences irrespective of the mdx mutation. Our study confirms a hierarchical, tripartite organisation of the skull (comprising neurocranium, face and mandible) with a modular division based on development and function. Moreover, we provide further experimental evidence that masticatory loading is one of the main environmental stimuli that generate craniofacial variation.

A dental perspective on the taxonomic affinity of the Balanica mandible (BH-1).

The Middle Pleistocene represents a period of critical importance in human evolution, marked by encephalisation and dental reduction, and increasing diversification of temporally and spatially distributed hominin lineages in Africa, Asia and Europe. New specimens, especially from areas less well represented in the fossil record, can inform the debate on morphological changes to the skeleton and teeth and the phylogenetic course of human evolution during this period. The mandible from the cave of Mala Balanica, Serbia has recently been re-dated to at least 400 ka, and its well-preserved dentition presents an excellent opportunity to characterize molar crown morphology at this time period, and re-examine claims for a lack of Neandertal affinities in the specimen. In this study we employ microtomography to image the internal structure of the mandibular molars (focusing on the morphology of the enamel-dentine junction, or EDJ) of the BH-1 specimen and a comparative sample (n = 141) of Homo erectus sensu lato, Homo neanderthalensis, Pleistocene Homo sapiens, and recent H. sapiens. We quantitatively assess EDJ morphology using 3D geometric morphometrics and examine the expression of discrete dental traits at the dentine surface. We also compare third molar enamel thickness in BH-1 to those of H. neanderthalensis and both Pleistocene and recent H. sapiens, and document previously unreported morphology of the BH-1 premolar and molar roots. Our results highlight the reliability of the EDJ surface for classifying hominin taxa, indicate a primitive dental morphology for BH-1 molars, and confirm a general lack of derived Neandertal features for the Balanica individual. The plesiomorphic character of BH-1 is consistent with several competing models of Middle Pleistocene hominin evolution and provides an important regional and temporal example for reconstructing morphological changes in the mandible and teeth during this time period.

Reconstruction of muscle fascicle architecture from iodine-enhanced microCT images: A combined texture mapping and streamline approach.

Skeletal muscle models are used to investigate motion and force generation in both biological and bioengineering research. Yet, they often lack a realistic representation of the muscle's internal architecture which is primarily composed of muscle fibre bundles, known as fascicles. Recently, it has been shown that fascicles can be resolved with micro-computed tomography (µCT) following staining of the muscle tissue with iodine potassium iodide (I2KI). Here, we present the reconstruction of the fascicular spatial arrangement and geometry of the superficial masseter muscle of a dog based on a combination of pattern recognition and streamline computation. A cadaveric head of a dog was incubated in I2KI and µCT-scanned. Following segmentation of the masseter muscle a statistical pattern recognition algorithm was applied to create a vector field of fascicle directions. Streamlines were then used to transform the vector field into a realistic muscle fascicle representation. The lengths of the reconstructed fascicles and the pennation angles in two planes (frontal and sagittal) were extracted and compared against a tracked fascicle field obtained through cadaver dissection. Both fascicle lengths and angles were found to vary substantially within the muscle confirming the complex and heterogeneous nature of skeletal muscle described by previous studies. While there were significant differences in the pennation angle between the experimentally derived and µCT-reconstructed data, there was congruence in the fascicle lengths. We conclude that the presented approach allows for embedding realistic fascicle information into finite element models of skeletal muscles to better understand the functioning of the musculoskeletal system.