Technical note: Virtual reconstruction of a fragmentary clavicle

We report a procedure for the virtual reconstruction of incomplete human bones applicable to skeletal remains from archaeological excavations or to reconstructive and prosthetic surgery. To test the procedure, we reconstructed a fragmented left clavicle on the basis of the contralateral clavicle. The procedure involved 3‐D laser scanner acquisition of the left clavicle (complete but broken into two parts), the same manually reconstructed bone, and the intact right clavicle, which was mirror‐imaged and used as a reference for the reconstruction of the whole left clavicle. Because it was not possible to recognize homologous anatomical landmarks, on the two reference models (a mirror‐image copy of the right clavicle and the main fragment of the left), we identified three grids with an increasing number of corresponding landmarks, which constituted the framework of the deformation process. The three reconstructed digital models of the clavicle closely approximated the model of the original clavicle. They also showed that an increasing number of landmarks did not significantly improve the reconstructed model. Am J Phys Anthropol 2009. © 2009 Wiley‐Liss, Inc.     

Chitosan‐heparin polyelectrolyte multilayers on cortical bone: Periosteum‐mimetic,cytophilic, antibacterial coatings

Cortical bone allografts suffer from high rates of failure due to poor integration with host tissue, leading to non‐union, fracture, and infection following secondary procedures. Here, we report a method for modifying the surfaces of cortical bone with coatings that have biological functions that may help overcome these challenges. These chitosan‐heparin coatings promote mesenchymal stem cell attachment and have significant antibacterial activity against both S. aureus and E. coli. Furthermore, their chemistry is similar to coatings we have reported on previously, which effectively stabilize and deliver heparin‐binding growth factors. These coatings have potential as synthetic periosteum for improving bone allograft outcomes. Biotechnol. Bioeng. 2013; 110: 609–618. © 2012 Wiley Periodicals, Inc.     

Longitudinal and lateral variations in the aluminum concentration of selected caprine, bovine, and human bone samples

Longitudinal and lateral variations in Al concentration in several large animal (bovine and caprine) long bones (tibia and femur) and several human clavicle bones were examined using a sensitive analytical method based on electrothermal atomization atomic absorption spectrometry with Zeeman background correction. Bone segments were carefully removed using special tools free of significant Al contamination, freeze-dried, and digested overnight at room temperature in concentrated HNO₃. Bone digestates were analyzed for Al using simple aqueous calibration standards with a Ca(NO₃)₂ modifier. Mean bone Al concentrations were relatively low (<1 Μg/g, dry weight) in bovine and caprine long bones compared to literature values for human bone samples. Longitudinal variations of Al in the animal bones examined appeared relatively uniform compared to the human clavicle bones, where, in three of five cases, Al appeared enriched at the epiphyses (joints). The Al “enrichment” was symmetrical with respect to both left and right clavicle bones. Aluminum concentrations at the mid-shaft of the clavicle bone show less variation compared to whole bone studies, but considerable scatter is evident along the bone length. The mean bone aluminum concentration in the five human subjects varied from 1 to 6 Μ/g dry weight.     

Tendon‐to‐bone attachment: From development to maturity

The attachment between tendon and bone occurs across a complex transitional tissue that minimizes stress concentrations and allows for load transfer between muscles and skeleton. This unique tissue cannot be reconstructed following injury, leading to high incidence of recurrent failure and stressing the need for new clinical approaches. This review describes the current understanding of the development and function of the attachment site between tendon and bone. The embryonic attachment unit, namely, the tip of the tendon and the bone eminence into which it is inserted, was recently shown to develop modularly from a unique population of Sox9‐ and Scx‐positive cells, which are distinct from tendon fibroblasts and chondrocytes. The fate and differentiation of these cells is regulated by transforming growth factor beta and bone morphogenetic protein signaling, respectively. Muscle loads are then necessary for the tissue to mature and mineralize. Mineralization of the attachment unit, which occurs postnatally at most sites, is largely controlled by an Indian hedgehog/parathyroid hormone‐related protein feedback loop. A number of fundamental questions regarding the development of this remarkable attachment system require further study. These relate to the signaling mechanism that facilitates the formation of an interface with a gradient of cellular and extracellular phenotypes, as well as to the interactions between tendon and bone at the point of attachment. Birth Defects Research (Part C) 102:101–112, 2014. © 2014 Wiley Periodicals, Inc.     

Evolution of hindlimb bone dimensions and muscle masses in house mice selectively bred for high voluntary wheel‐running behavior

We have used selective breeding with house mice to study coadaptation of morphology and physiology with the evolution of high daily levels of voluntary exercise. Here, we compared hindlimb bones and muscle masses from the 11th generation of four replicate High Runner (HR) lines of house mice bred for high levels of voluntary wheel running with four non‐selected control (C) lines. Mass, length, diameter, and depth of the femur, tibia‐fibula, and metatarsal bones, as well as masses of gastrocnemius and quadriceps muscles, were compared by analysis of covariance with body mass or body length as the covariate. Mice from HR lines had relatively wider distal femora and deeper proximal tibiae, suggesting larger knee surface areas, and larger femoral heads. Sex differences in bone dimensions were also evident, with males having thicker and shorter hindlimb bones when compared with females. Several interactions between sex, linetype, and/or body mass were observed, and analyses split by sex revealed several cases of sex‐specific responses to selection. A subset of the HR mice in two of the four HR lines expressed the mini‐muscle phenotype, characterized mainly by an ～50% reduction in hindlimb muscle mass, caused by a Mendelian recessive mutation, and known to have been under positive selection in the HR lines. Mini‐muscle individuals had elongated distal elements, lighter and thinner hindlimb bones, altered 3rd trochanter muscle insertion positions, and thicker tibia‐fibula distal widths. Finally, several differences in levels of directional or fluctuating asymmetry in bone dimensions were observed between HR and C, mini‐ and normal‐muscled mice, and the sexes. This study demonstrates that skeletal dimensions and muscle masses can evolve rapidly in response to directional selection on locomotor behavior.     

Male sexual ornament size but not asymmetry reflects condition in stalk-eyed flies

Models of sexual selection predict that females use ornament size to evaluate male condition. It has also been suggested that ornament asymmetry provides females with accurate information about condition. To test these ideas we experimentally manipulated condition in the stalk-eyed fly, Cyrtodiopsis dalmanni, by varying the amount of food available to developing larvae. Males of this species have greatly exaggerated eyestalk length and females prefer to mate with males with wider eyespans. Our experiments show that male ornaments (eyestalks) display a disproportionate sensitivity to condition compared with the homologous character in females, and to non-sexual traits (wing dimensions). In contrast, in neither sex did asymmetry reflect condition either in sexual ornaments or in non-sexual traits. We conclude that ornament size is likely to play a far greater role in sexual selection as an indicator of individual condition than does asymmetry.     

FLUCTUATING ODONTOMETRIC ASYMMETRY,MORPHOLOGICAL VARIABILITY,AND GENETIC MONOMORPHISM IN THE CHEETAH ACINONYX JUBATUS

The magnitudes of dimensional variability and fluctuating asymmetry in dental dimensions are reported for a sample of South African cheetah Acinonyx jubatus. To test the hypothesis that elevated levels of variability and asymmetry are associated with the increased developmental instability reported for this species, our results were contrasted to those for two other felids: Felis lybica and F. caracal. These findings suggest that dental dimensions in cheetahs are not significantly more variable or asymmetric. Hence, it is concluded that the cheetah may not be as developmentally unstable as was previously supposed.     

External-internal relationships in humeri of modern human females

With data from an early twentieth century human skeletal collection, this exploratory study investigates associations between inner cortical and medullary cavity structures and outer shaft and epiphyseal features of a long bone. Humeri are measured directly in both whole bone and transverse section contexts; data along 2 axes at 2 sites are obtained. Twenty-two probable females, with an age range concentrated in middle adulthood, comprise the sample. Correlations between multiple external and internal bone measurements are analyzed, with the aim of yielding information on the physical nature of bone and on the effects of different measurement types, locations, and orientations for bone relationships. The study also examines whether prediction of inner humeral dimensions from outer measurements would be appropriate. Results indicate biepicondylar width and maximum length as the strongest external correlates of cortical dimensions. Contrasting with studies on the proximal femur, the humeral head shows external size changeability, mostly in the transverse plane, in response to modeling forces shared with the shaft. Epiphyseal measures are more highly associated with absolute rather than percent, and areal rather than linear, cortical variables. Medullary cavity dimensions are not significantly correlated with whole bone measures. Most associations demonstrate shape or proportion influences rather than a shared effect of linear body size. Regarding location and orientation, the distal site and medial-lateral axis display the strongest correlations among external and internal bone dimensions. In light of the demonstrated correlation patterns, prediction of humeral cortical quantity from external bone measures in living and skeletal populations would benefit from utilizing (1) biepicondylar width and maximum length; (2) an absolute, areal cortical measure; (3) a distal location; and (4) a medial-lateral orientation.     

Age estimation of adolescent and young adult male and female skeletons II, epiphyseal union at the upper limb and scapular girdle in a modern Portuguese skeletal sample

This study completes previously reported ages for timing of epiphyseal union in the postcranial skeleton in a recent sample, with data from the scapula, clavicle, humerus, radius, and ulna. A sample of 121 individuals between the ages of 9 and 29 (females = 65, males = 56) was derived from the Lisbon documented skeletal collection. Epiphyseal union was scored at 16 anatomical locations, using a three-stage scheme: 1) no union; 2) partial union; and 3) completed union, all traces of fusion having disappeared. In the upper limb, the epiphyses of the elbow are the first to fuse at around 11 to 15 years of age, followed by those of the shoulder and wrist. In the scapular girdle, the coracoid area is the first to fuse, followed by the glenoid surface and remaining epiphyses, with the medial clavicle fusing last, by the age of 25-27. There is a sex difference in maturation, with females showing an advance relative to males of about 2 years in the upper limb. Sex differences in maturation are less noticeable in the scapular girdle, but data suggest that females are also ahead of males. Results suggest overall similar age ranges for stages of union as previous dry bone observations, but some studies show significant divergences which seem to derive from methodological issues. Although some radiographic reference standards provide comparable age ranges, they should probably be avoided when aging skeletal remains.     

Tracing prehistoric activities: musculoskeletal stress marker analysis of a Stone-Age population on the island of Gotland in the Baltic sea

The skeletal remains from the Middle Neolithic (2750-2300 BC) burial ground at Ajvide, Gotland, are analyzed in order to explore musculoskeletal patterns and to attempt to trace general as well as three specific prehistoric activities (archery, harpooning, and kayaking) that are likely to have been performed in this marine setting of fishing, hunting, and gathering. Scoring of muscular and ligament attachments is performed using the scoring method of Hawkey and Merbs ([1995] Int. J. Osteoarchaeol. 5:324-338) for muskuloskeletal stress markers (MSM). The skeletal material consists of 24 male and 15 female adult individuals divided into three age groups: young (<24 years), middle (25-39 years), and old (>40 years). Thirty upper body MSM sites, on both the left and right sides, are scored and form the basis of the study. Results show that males most frequently have higher mean MSM scores than females. Bilateral asymmetry was noted as low in both sexes. Age proved to be a contributing factor to increased MSM scores, with a greater age-related increase in females. MSM patterns were analyzed statistically in muscle groups associated with the three investigated activities. Significant positive correlations were observed in male individuals in muscle groups associated with archery and to some extent harpooning, an indication that these activities would mainly have been performed by men. Correlations in kayaking muscles were not evidently consistent with the kayaking motion. Furthermore, the costoclavicular ligament, often referred to in connection with "kayaker's clavicle," showed no positive statistical correlation with the kayaking muscles.     

Cortical bone thickness can adapt locally to muscular loading while changing with age

Mechanical loading of muscle action is concentrated at muscle attachment sites; thus there may be a potential for site-specific variation in cortical bone thickness. Humeri from an early 20th-century Finnish (Helsinki) and two medieval English (Newcastle, Blackgate and York, Barbican) populations were subjected to pQCT scanning to calculate site-specific cross-sectional cortical bone area (CA) for four locations and to measure cortical thickness at muscle attachment sites and non-attachment sites. We found that CA at 80% of humerus length was significantly reduced compared to more distal cross-sections, which can be due to reduced stresses at the proximal shaft. The principal direction of loading at 80% humerus length was towards mediolateral plane, likely due to fixing the humerus close to the torso. At 35% the main direction of loading was towards anteroposterior plane, reflecting elbow flexing forces. The principal direction of loading varied between populations, sides and sexes at 50% humerus length due to preference between elbow and shoulder joint; thus this location might be useful when trying to infer differences in activity. These changes are likely due to overall shaft adaptation to forces acting at the humerus. In addition, we found a potential for site-specific variation in cortical thickness; cortical bone at muscle attachment sites was significantly thicker compared to non-attachment sites. Lastly, CA at 35% of humerus length and cortical thickness at non-attachment sites decreased with age. These results underline the importance of muscle loading for bone mass preservation as well as indicate that a site-specific variation of bone mass is possible.     

The importance of accounting for the area of the medullary cavity in cross‐sectional geometry: A test based on the femoral midshaft

In cross‐sectional geometric (CSG) studies, both the subperiosteal and endosteal contours are considered important factors in determining bone bending rigidity. Recently, regression equations predicting CSG properties from a section's external dimensions were developed in a world‐wide sample of human long bones. The results showed high correlations between some subperiosteally derived and actual CSG parameters. We present a theoretical model that further explores the influence of endosteal dimensions on CSG properties. We compare two hypothetical femoral midshaft samples with the same total subperiosteal area but with percentages of cortical bone at the opposite ends of published human variation for population sample means. Even in this relatively uncommon scenario, the difference between the samples in the resultant means for predicted femoral polar second moment of area (J) appears to be modest: power analysis indicates that a minimum sample size of 61 is needed to detect the difference 90% of the time via a t‐test. Moreover, endosteal area can be predicted—although with substantial error—from periosteal area. Despite this error, including this relationship in subperiosteally derived estimates of J produces sample mean estimates close to true mean values. Power analyses reveal that when similar samples are used to develop prediction equations, a minimum sample of hundreds or more may be needed to distinguish a predicted mean J from the true mean J. These results further justify the use of regression equations estimating J from periosteal contours when analyzing behaviorally induced changes in bone rigidity in ancient populations, when it is not possible to measure endosteal dimensions. However, in other situations involving comparisons of individual values, growth trends, and senescence, where relative cortical thickness may vary greatly, inclusion of endosteal dimensions is still important. Am J Phys Anthropol, 2010. © 2010 Wiley‐Liss, Inc.     

Estimating the length of incomplete long bones: forensic standards from Guatemala

We report on new standards for estimating long bone length from incomplete bones for use in forensic and archaeological contexts in Central America. The measurements we use closely follow those defined by Steele ([1970] Personal Identification in Mass Disasters; Washington, DC: Smithsonian Institution), but we add several new landmarks. We measured the femur, humerus, tibia, and fibula of 100 Maya skeletons (68 males, 32 females) recovered from forensic exhumations. We derived the equations by regressing bone segment length on bone length, and solved for bone length to maximize the utility of the equations for taller populations. We generated equations for all segments that were significantly correlated with bone length for males, for females, and for both sexes combined, but accepted only regressions with r(2) > 0.85 as reliable. Landmarks defined by muscle attachment sites were more variable in location than landmarks on articular architecture; thus we retained few equations that use these landmarks. We tested the male and combined sex equations on 36 males of unknown ethnicity exhumed from a military base in Guatemala, and found that the equations performed satisfactorily. We also evaluated the performance of equations by Steele ([1970] Personal Identification in Mass Disasters; Washington, DC: Smithsonian Institution) and Jacobs ([1992] Am J Phys Anthropol 89:333-345) on the Maya bones, and conclude that significant population variation in long bone proportions hinders their application in Central America.     

Left‐right asymmetry in gut development: what happens next?

The gastrointestinal tract is an asymmetrically patterned organ system. The signals which initiate left‐right asymmetry in the developing embryo have been extensively studied, but the downstream steps required to confer asymmetric morphogenesis on the gut organ primordia are less well understood. In this paper we outline key findings on the tissue mechanics underlying gut asymmetry, across a range of species, and use these to synthesise a conserved model for asymmetric gut morphogenesis. We also discuss the importance of correct establishment of left‐right asymmetry for gut development and the consequences of perturbations in this process.     

Evaluation of a morphing based method to estimate muscle attachment sites of the lower extremity

To generate subject-specific musculoskeletal models for clinical use, the location of muscle attachment sites needs to be estimated with accurate, fast and preferably automated tools. For this purpose, an automatic method was used to estimate the muscle attachment sites of the lower extremity, based on the assumption of a relation between the bone geometry and the location of muscle attachment sites. The aim of this study was to evaluate the accuracy of this morphing based method. Two cadaver dissections were performed to measure the contours of 72 muscle attachment sites on the pelvis, femur, tibia and calcaneus. The geometry of the bones including the muscle attachment sites was morphed from one cadaver to the other and vice versa. For 69% of the muscle attachment sites, the mean distance between the measured and morphed muscle attachment sites was smaller than 15 mm. Furthermore, the muscle attachment sites that had relatively large distances had shown low sensitivity to these deviations. Therefore, this morphing based method is a promising tool for estimating subject-specific muscle attachment sites in the lower extremity in a fast and automated manner.     

Directional asymmetry of body dimensions among white adolescents

Asymmetry of paired dimensions has been recognized as a methodological problem in anthropometry and more recently as an indicator of environmental stress. This study seeks to determine the extent of directional asymmetry for some of the measurements commonly made in anthropometry. Upper arm circumference, biepicondylar breadth, triceps and subscapular skinfolds, bicondylar breadth of the femur, and calf circumference were measured on right and left sides among 135 white adolescents from suburban Philadelphia. Handedness (right or nonright) was subject-assessed. Body composition was estimated through underwater weighing. Asymmetry was evaluated using a paired t test. Arm measurements are significantly asymmetric in favor of the right side; subscapular skinfolds and leg measurements are not significantly asymmetric. Among the sample of right-handed subjects (n = 116), upper arm circumference and biepicondylar breadth were significantly larger on the right side, and, among the males of this subsample, triceps was as well. The nonright-handed subjects (n = 19) did not show statistically significant asymmetry. Asymmetry was negatively but weakly related to body composition. These results are consistent with an explanation in terms of preferred use of one side of the body and consequent muscle hypertrophy, but an adequate test of this explanation requires hypothesis testing in larger samples of nonright-handed subjects.     

Development of an anatomically based whole‐body musculoskeletal model of the Japanese macaque (Macaca fuscata)

We constructed a three‐dimensional whole‐body musculoskeletal model of the Japanese macaque (Macaca fuscata) based on computed tomography and dissection of a cadaver. The skeleton was modeled as a chain of 20 bone segments connected by joints. Joint centers and rotational axes were estimated by joint morphology based on joint surface approximation using a quadric function. The path of each muscle was defined by a line segment connecting origin to insertion through an intermediary point if necessary. Mass and fascicle length of each were systematically recorded to calculate physiological cross‐sectional area to estimate the capacity of each muscle to generate force. Using this anatomically accurate model, muscle moment arms and force vectors generated by individual limb muscles at the foot and hand were calculated to computationally predict muscle functions. Furthermore, three‐dimensional whole‐body musculoskeletal kinematics of the Japanese macaque was reconstructed from ordinary video sequences based on this model and a model‐based matching technique. The results showed that the proposed model can successfully reconstruct and visualize anatomically reasonable, natural musculoskeletal motion of the Japanese macaque during quadrupedal/bipedal locomotion, demonstrating the validity and efficacy of the constructed musculoskeletal model. The present biologically relevant model may serve as a useful tool for comprehensive understanding of the design principles of the musculoskeletal system and the control mechanisms for locomotion in the Japanese macaque and other primates. Am J Phys Anthropol, 2009. © 2008 Wiley‐Liss, Inc.     

Radiographic changes in the clavicle and proximal femur and their use in the determination of skeletal age at death

Visually seriated radiographs of the proximal femur, proximal humerus, clavicle, and calcaneus from 130 individuals from the Hamann-Todd collection were examined as indicators of skeletal age at death. The clavicle demonstrated the most consistent relationship to age in both sexes. The same radiographs were also seriated by size-normalized optical density as a means of establishing relative radiolucency. In this context, visual seriation proved superior. The four sites studied showed strong divergence in response to age. Since each was sampling bone response from the same individual, it is concluded that bone loss is highly site specific. This demonstrates the individual character of specific skeletal sites. Visual inspection of clavicular radiographs, seriated on a populational basis, provides age estimates that are comparable to anatomical age indicators and provides independent estimates of skeletal age when included in the summary age method (1985: Am. J. Phys. Anthropol. 68:1–14).     

Dimorphism of right–left asymmetry in the female genitalia of the brachypterous grasshopper Parapodisma mikado (Orthoptera: Acrididae)

Several insects exhibit morphological asymmetry in the mouthparts or genitalia. In a part of species with asymmetric genitalia, two mirror‐image forms of the genitalia are reported to occur in a population. This dimorphism, called chiral dimorphism, is usually observed in male genitalia, but its examples in female genitalia are very limited. Here, we report that the females of the brachypterous grasshopper Parapodisma mikado are equipped with an asymmetric sclerite in the copulatory bursa, and that two mirror‐image forms of the sclerite occur in local populations. This dimorphism was detected in all of seven populations examined, and the ratio of the two forms was approximately 2:1 for the right : left forms. In one population, the ratio of the two forms did not vary among three consecutive years. Thus, chiral dimorphism in the female genitalia is kept almost constant spatiotemporally. Mating experiments indicated that the direction of female genitalia did not affect the direction to which the males twist their abdomen (right or left side) during mating or the duration of mating. We propose two hypotheses, a neutral developmental mechanism and sexual conflict, for the maintenance of chiral dimorphism in the genitalia of P. mikado females.     

Mapping discontinuous protein‐binding sites via structure‐based peptide libraries: combining in silico and in vitro approaches

To perform their various functions, protein surfaces often have to interact with each other in a specific way. Usually, only parts of a protein are accessible and can act as binding sites. Because proteins consist of polypeptide chains that fold into complex three‐dimensional shapes, binding sites can be divided into two different types: linear sites that follow the primary amino acid sequence and discontinuous binding sites, which are made up of short peptide fragments that are adjacent in spatial proximity. Such discontinuous binding sites dominate protein–protein interactions, but are difficult to identify. To meet this challenge, we combined a computational, structure‐based approach and an experimental, high‐throughput method. SUPERFICIAL is a program that uses protein structures as input and generates peptide libraries to represent the protein's surface. A large number of the predicted peptides can be simultaneously synthesised applying the SPOT technology. The results of a binding assay subsequently help to elucidate protein–protein interactions; the approach is applicable to any kind of protein. The crystal structure of the complex of hen egg lysozyme with the well‐characterised murine IgG1 antibody HyHEL‐5 is available, and the complex is known to have a discontinuous binding site. Using SUPERFICIAL, the entire surface of lysozyme was translated into a peptide library that was synthesised on a cellulose membrane using the SPOT technology and tested against the HyHEL‐5 antibody. In this way, it was possible to identify two peptides (longest common sequence and peptide 19) that represented the discontinuous epitope of lysozyme. Copyright © 2012 John Wiley & Sons, Ltd.