Comparing rates of recrystallisation and the potential for preservation of biomolecules from the distribution of trace elements in fossil bones

Preservation of intact macromolecules and geochemical signals in fossil bones is mainly controlled by the extent of post-mortem interaction between bones and sediment pore waters. Trace elements such as lanthanum are added to bone post-mortem from pore waters, and where uptake occurs via a simple process of diffusion and adsorption, the elemental distribution can be used to assess the relative extent of bone-pore water interaction and rate of recrystallisation. Distribution profiles can be parameterised effectively using simple exponential equations, and the extent of bone–water interaction compared within and between sites. In this study, the distribution of lanthanum within bone was determined by laser ablation ICP–MS in 60 archaeological and fossil bones from Pleistocene and Cretaceous sites. The rates of recrystallisation and potential for preservation of intact biogeochemical signals vary significantly within and between sites. Elemental profiles within fossil bones hold promise as a screening technique to prospect for intact biomolecules and as a taphonomic tool.     

Effects of thirty elements on bone metabolism

The human skeleton, made of 206 bones, plays vital roles including supporting the body, protecting organs, enabling movement, and storing minerals. Bones are made of organic structures, intimately connected with an inorganic matrix produced by bone cells. Many elements are ubiquitous in our environment, and many impact bone metabolism. Most elements have antagonistic actions depending on concentration. Indeed, some elements are essential, others are deleterious, and many can be both. Several pathways mediate effects of element deficiencies or excesses on bone metabolism. This paper aims to identify all elements that impact bone health and explore the mechanisms by which they act. To date, this is the first time that the effects of thirty minerals on bone metabolism have been summarized.     

Bone protects proteins over thousands of years: extraction, analysis, and interpretation of extracellular matrix proteins in archeological skeletal remains

In a good state of preservation, bone conserves the entire protein pattern of extracellular bone matrix proteins over thousands of years. The quality of the profiles of matrix proteins isolated from ancient bones (ranging from the pre-Pottery Neolithic Phase to Early Modern Times from different archaeological sites in different geographical areas), separated by electrophoresis, is as good as those from recent bones. Molecules arising from collagenous proteins (e.g., collagen type I), from the noncollagenous group (e.g., osteonectin), and from the immune system (e.g., immunoglobulin G) were identified in Western blots by specific antibodies. A comparative study of the immunoglobulin G content of the bones of five prehistoric children showed the lowest immunoglobulin G content in a child who suffered from chronic scurvy. Ancient bone proteins were also separated by two-dimensional electrophoresis. This technique makes fractionation of the complex protein mixtures of extracellular bone matrix more reproducible. Bone retains a chemical memory of earlier metabolic stimuli in its configuration of collagenous and noncollagenous proteins. In combination with the results of the microscopic examination of ancient bone, it should be possible to obtain more reliable information on the history and the evolution of diseases, based on analysis of intact proteins.     

Investigating histomorphological variations in human cranial bones through ontogeny

Through ontogeny, human cranial vault bones undergo differentiation in terms of their shape, size and tissue maturation. This differentiation is visible at both the macroscopic and microscopic levels. Preliminary data from a histological and compartmentalisation exploratory analysis of individuals with different ages suggest differences in the modelling and remodelling patterns through ontogeny. Child vault bones are primarily composed of avascular lamellar bone (largely vascularised), late juvenile or adolescent bones present the largest extension of mineralised areas (highly remodelled) and the lowest vascularisation (diploe is highly reduced), and the adult present highly vascularised bone in which the diploe is again largely extended. During childhood, the existence of an avascular lamellar bone promotes the sealing of the cranium bones surfaces whereas adult vault bones seem to become opened ectocranially due to the remodelling. We discuss the possibility that both effects could be related with the head thermoregulation.     

Ca/P concentration ratio at different sites of normal and osteoporotic rabbit bones evaluated by Auger and energy dispersive X-ray spectroscopy

Osteoporosis is a systemic skeletal disorder associated with reduced bone mineral density and the consequent high risk of bone fractures. Current practice relates osteoporosis largely with absolute mass loss. The assessment of variations in chemical composition in terms of the main elements comprising the bone mineral and its effect on the bone’s quality is usually neglected. In this study, we evaluate the ratio of the main elements of bone mineral, calcium (Ca), and phosphorus (P), as a suitable in vitro biomarker for induced osteoporosis. The Ca/P concentration ratio was measured at different sites of normal and osteoporotic rabbit bones using two spectroscopic techniques: Auger electron spectroscopy (AES) and energy-dispersive X-ray spectroscopy (EDX). Results showed that there is no significant difference between samples from different genders or among cortical bone sites. On the contrary, we found that the Ca/P ratio of trabecular bone sections is comparable to cortical sections with induced osteoporosis. Ca/P ratio values are positively related to induced bone loss; furthermore, a different degree of correlation between Ca and P in cortical and trabecular bone is evident. This study also discusses the applicability of AES and EDX to the semiquantitative measurements of bone mineral’s main elements along with the critical experimental parameters.     

Distribution of trace elements within bones in nasal cavity and labyrinth in humans

Measurements of trace element concentrations within bones in nasal cavity and labyrinth have shown large variations, both with a single bone and between different bones of a same individual. Factors that influence trace element levels include: metabolic activity, environmental effects, sex, and age. Detection of characteristic X-rays has been shown to be a convenient method for the measurement of concentration profiles, micropixe for micrometer variations, and X-ray centration profiles, micropixe for micrometer variations, and X-ray fluorescence for millimeter variations.     

Taphonomic Analysis of the Faunal Assemblage Associated with the Hominins (Australopithecus sediba) from the Early Pleistocene Cave Deposits of Malapa,South Africa

Here we present the results of a taphonomic study of the faunal assemblage associated with the hominin fossils (Australopithecus sediba) from the Malapa site. Results include estimation of body part representation, mortality profiles, type of fragmentation, identification of breakage patterns, and microscopic analysis of bone surfaces. The diversity of the faunal spectrum, presence of animals with climbing proclivities, abundance of complete and/or articulated specimens, occurrence of antimeric sets of elements, and lack of carnivore-modified bones, indicate that animals accumulated via a natural death trap leading to an area of the cave system with no access to mammalian scavengers. The co-occurrence of well preserved fossils, carnivore coprolites, deciduous teeth of brown hyaena, and some highly fragmented and poorly preserved remains supports the hypothesis of a mixing of sediments coming from distinct chambers, which collected at the bottom of the cave system through the action of periodic water flow. This combination of taphonomic features explains the remarkable state of preservation of the hominin fossils as well as some of the associated faunal material.     

The Exploitation of Ungulate Bones in Homo neanderthalensis and Homo sapiens

Analysis of ungulate bones recovered from a number of Upper and Middle Palaeolithic sites in southern Italy revealed differences in the presence of anatomical elements. There is a lack of clear evidence of carnivore activities, and differences can be attributed to human activity. Indeed, these differences were probably due to different patterns of skeletal exploitation between Homo neanderthalensis and H. sapiens. Small limb bones (carpals, tarsals, sesamoids, long bone epiphyses and especially phalanges) are rarely found in Middle Palaeolithic deposits, but are abundant in the Upper Palaeolithic. The observation of unidentified bone fragments at these sites indicates that during the middle Palaeolithic, marrow extraction regarded essentially the treatment of long bones. First and second phalanges were not frequently used for this practice, but they were often fragmented by H. sapiens. Lack of these bones among the remains of meals of Neanderthal suggests that these bones were probably destroyed by their utilisation as fuel.     

Embryonic development of Python sebae - I: Staging criteria and macroscopic skeletal morphogenesis of the head and limbs

This study explores the post-ovipositional craniofacial development of the African Rock Python (Python sebae). We first describe a staging system based on external characteristics and next use whole-mount skeletal staining supplemented with Computed tomography (CT) scanning to examine skeletal development. Our results show that python embryos are in early stages of organogenesis at the time of laying, with separate facial prominences and pharyngeal clefts still visible. Limb buds are also visible. By 11 days (stage 3), the chondrocranium is nearly fully formed; however, few intramembranous bones can be detected. One week later (stage 4), many of the intramembranous upper and lower jaw bones are visible but the calvaria are not present. Skeletal elements in the limbs also begin to form. Between stages 4 (day 18) and 7 (day 44), the complete set of intramembranous bones in the jaws and calvaria develops. Hindlimb development does not progress beyond stage 6 (33 days) and remains rudimentary throughout adult life. In contrast to other reptiles, there are two rows of teeth in the upper jaw. The outer tooth row is attached to the maxillary and premaxillary bones, whereas the inner row is attached to the pterygoid and palatine bones. Erupted teeth can be seen in whole-mount stage 10 specimens and are present in an unerupted, mineralized state at stage 7. Micro-CT analysis reveals that all the young membranous bones can be recognized even out of the context of the skull. These data demonstrate intrinsic patterning of the intramembranous bones, even though they form without a cartilaginous template. In addition, intramembranous bone morphology is established prior to muscle function, which can influence bone shape through differential force application. After careful staging, we conclude that python skeletal development occurs slowly enough to observe in good detail the early stages of craniofacial skeletogenesis. Thus, reptilian animal models will offer unique opportunities for understanding the early influences that contribute to perinatal bone shape.     

安徽琅琊山铜矿蝶类重元素X荧光分析

为了研究安徽琅琊山铜矿环境中重元素污染的状况,利用同步辐射X射线荧光,对于采自安徽安徽琅琊山铜矿的9种蝴蝶的翅样品进行重元素分析。结果显示,9种蝴蝶体内重元素含量的差别变化明显,蝴蝶可以富集重元素Hg和Pb,其Cu、Zn元素的比值与环境污染程度相关。结果表明该方法样品处理简便,灵敏度高。     

Allometry of cetacean forelimb bones

This study examines the allometric scaling relationships of the cetacean humerus, radius, and ulna. Bone lengths and diameters were measured for 20 species of odontocete and three species of mysticete cetaceans, representing eight of the nine extant cetacean families. The scaling of individual bone proportions (bone length vs. cranio-caudal diameter, bone length vs. dorso-ventral diameter), and of individual bone dimensions against estimated body mass, are compared to models of geometric and elastic similarity. The geometric similarity model describes the scaling relationship of bone length vs. cranio-caudal diameter and body mass vs. cranio-caudal diameter for the humerus only; geometric similarity also describes the scaling relationship of body mass vs. bone length for all three bones. None of the scaling relationships fits the elastic similarity model. The scaling relationships of bone length vs. dorso-ventral diameter for all three bones, and bone length vs. cranio-caudal diameter for the radius and ulna, exhibit negative allometry, indicating that large bones are less robust than small bones. Negative allometry of structural support elements has not been previously described for terrestrial mammals or plants. The high relative swimming speeds of small delphinids may generate sufficient stresses to require more robust bones relative to those of larger whales. © 1994 Wiley-Liss, Inc.     

Concentrations of trace elements in bones of the hip joint from patients after hip replacement surgery

The aim of this research was to determine the concentrations of two essential elements (copper (Cu) and zinc (Zn)) and three toxic elements (lead (Pb), cadmium (Cd) and mercury (Hg)) in the hip joint bones of patients from the Chair and Clinic of Orthopaedics and Traumatology at the Pomeranian Medical University in Szczecin. We examined 111 samples of hip joint bones obtained from patients from north-western Poland who had undergone arthroplasty of the femoral head. In cartilage with the adjacent compact bone, and in spongy bone from the examined patients, the concentrations (medians) determined were placed in the following descending sequence Zn>Pb>Cu>Cd>Hg. The concentrations ranged from 86 mg Zn/kg to 0.0020 mg Hg/kg dw. It was found that the concentration of lead in the cartilage with adjacent compact bone was higher in men than in women. In conclusion, it seems that in addition to routine monitoring of the abiotic environment, it is essential to monitor concentrations of heavy metals having a long-term impact in humans.     

Ancient pathogen DNA in human teeth and petrous bones

Recent ancient DNA (aDNA) studies of human pathogens have provided invaluable insights into their evolutionary history and prevalence in space and time. Most of these studies were based on DNA extracted from teeth or postcranial bones. In contrast, no pathogen DNA has been reported from the petrous bone which has become the most desired skeletal element in ancient DNA research due to its high endogenous DNA content. To compare the potential for pathogenic aDNA retrieval from teeth and petrous bones, we sampled these elements from five ancient skeletons, previously shown to be carrying Yersinia pestis. Based on shotgun sequencing data, four of these five plague victims showed clearly detectable levels of Y. pestis DNA in the teeth, whereas all the petrous bones failed to produce Y. pestis DNA above baseline levels. A broader comparative metagenomic analysis of teeth and petrous bones from 10 historical skeletons corroborated these results, showing a much higher microbial diversity in teeth than petrous bones, including pathogenic and oral microbial taxa. Our results imply that although petrous bones are highly valuable for ancient genomic analyses as an excellent source of endogenous DNA, the metagenomic potential of these dense skeletal elements is highly limited. This trade‐off must be considered when designing the sampling strategy for an aDNA project.     

The development of acellularity of the vertebral bone of the Japanese medaka,Oryzias latipes (Teleostei; Cyprinidontidae)

Among vertebrates, some teleosts are unique in having bone which lacks osteocytes embedded in the matrix. The fate of cells that secrete the matrix of these acellular bones has not been investigated thoroughly. Histological and fluorescent microscopic analysis of the vertebral bone of Oryzias latipes demonstrated that acellularity is not a secondary appearance of an early cellular bone during ontogeny. Vertebral bone is devoid of cells embedded in the matrix throughout development. Cells that secrete bone matrix do not become trapped in their own secretion. Instead, they always remain as a surface layer over the outer surface of the bone. Fluorescent microscopic visualization of tetracycline injected into growing fish demonstrated that bone was only deposited by osteoblasts lining the outer surface of the bone; no deposition of bone took place on the inner surface.     

Development of the head skeleton and pectoral girdle in Esox.

A consideration of head development in two species of Esox, lucius and americanus (ssp. vermiculatus) representing the two subgenera Esox and Kenozoa respectively, focused on the significance of the variations of the latero-sensory canal system, its associated bones, and other skeletal elements. In living forms only aspects of "regression" or specialization can be studied. Canals tend to be reduced to pit lines first at their termini but can be broken in their course. Pit lines range from nearly canals to surface structures, or even fail to develop. The number of neuromasts varies. Canal bones develop from two centers: neuromast related and deeper membranous centers which may have no relationship to neuromasts. Tooth-bearing and non-canal-related dermal bones have only membranous (original) centers. The number of neuromasts associated with a bone usually does not affect its development or form. In the case of the circumorbital bones, the extrascapulars, and the nasal, a one to one relationship has developed by regression--towards the development of the latero-sensory component only. The idea that reductions in bone number are commonly traceable to fusion is rejected although examples of fusion are know. Most bones that disappear are simply lost (no blastema or other evidence of their presence seen in development). The relationship between dermal bone and chondral bone is examined and there is evidence of the former giving rise to the latter. The ontogenic order of appearances shows a feeding (functional) correlation.     

Evaluation of Dog Bones in the Indirect Assessment of Environmental Contamination with Trace Elements

The aim of this paper was to determine the level of five elements, two essential for life [zinc (Zn) and copper (Cu)] and three distinctly toxic [lead (Pb), cadmium (Cd), and mercury (Hg)], in four types of biological material in bones of the dog Canis lupus familiaris. The experiment was carried out on bones from the hip joints of dogs. The samples of cartilage, compact bone, spongy bone, and cartilage with adjacent compact bone came from 26 domestic dogs from northwestern Poland. Concentrations of Cu, Zn, Pb, and Cd were determined by ICP-AES (atomic absorption spectrophotometry) in inductively coupled argon plasma, using a Perkin-Elmer Optima 2000 DV. Determination of Hg concentration was performed by atomic absorption spectroscopy. In the examined bone material from the dog, the greatest concentrations (median) were observed for Zn and the lowest for Hg (98 mg Zn/kg and 0.0015 mg Hg/kg dw, respectively). In cartilage and spongy bone, metal concentrations could be arranged in the following descending order: Zn > Pb > Cu > Cd > Hg. In compact bone, the order was slightly different: Zn > Pb > Cd > Cu > Hg (from median 70 mg/kg dw to 0.002 mg/kg dw). The comparisons of metal concentrations between the examined bone materials showed distinct differences only in relation to Hg: between concentrations in spongy bone, compact bone, and in cartilage, being greater in cartilage than in compact bone, and lower again in spongy bone.     

Osteohistological insight into the early stages of growth in Mussaurus patagonicus (Dinosauria,Sauropodomorpha)

Here, we describe the bone histology of juvenile specimens of the basal sauropodomorph Mussaurus patagonicus and interpret its significance in terms of the early growth dynamics of this taxon. Thin sections from three juvenile specimens (femur length, 111–120 mm) of Mussaurus were analysed. The sampled bones consist of multiple postcranial elements collected from the Late Triassic Laguna Colorada Formation (El Tranquilo Group, Patagonia). The cortical bone is composed of fibrolamellar bone tissue. Vascularisation is commonly laminar or plexiform in the long bones. Growth marks are absent in all the examined samples. The ‘epiphyses’ of long bones are all formed by well-developed hypertrophied calcified cartilage. The predominance of woven-fibred bone matrix in cortical bones indicates a fast growth rate in the individuals examined. Moreover, given the existence of growth marks in adult specimens of Mussaurus, as in other sauropodomorphs, and assuming that the first lines of arrested growth was formed during the first year of life, the absence of growth marks in all the bones suggest that the specimens died before reaching their first year of life. Compared with the African taxon Massospondylus carinatus (another basal sauropodomorph for which the bone histology has been previously studied), it appears that Mussaurus had a higher early growth rate than Massospondylus.     

Mineralogical–geochemical characteristics of the bone detritus of Pleistocene mammals as a source of paleontological information

A complex mineralogical geochemical study of the bone detritus of large Pleistocene mammals inhabiting the Pechora Fore-Urals, the northernmost European part of their geographical range, is performed for the first time. The chemical composition, microstructure, content of 50 chemical elements are analyzed. The taphonomy of bones, extent of preservation of their initial properties, character and extent of fossilization are considered. The ultraporosity in the nanometer range is examined for the first time; it is established that it can be a marker of epigenetic changes in bones, beginning from the earliest stage of their fossilization. Based on X-ray structure and spectroscopic data, bone bioapatite is referred to carbonate–apatite of the B type. The results of thermal, chromatographic, and spectroscopic studies of collagen extracted from the bones of Pleistocene animals are discussed for the first time. The general trend of chemical degradation of bone proteins during fossilization is determined. The amino acid composition of bone collagen, the isotope composition of C, O, and N in biomineral and organic components of bones in the course of two-stage mass-spectrometer analysis of a complex sample are analyzed for the first time. Based on isotope-geochemical data conclusions concerning the diet and paleoclimatic and paleolandscape conditions of Pleistocene mammals are made.     

Development and evolution of craniofacial patterning is mediated by eye-dependent and -independent processes in the cavefish Astyanax

We studied the development and evolution of craniofacial features in the teleost fish, Astyanax mexicanus. This species has an eyed surface dwelling form (surface fish) and many different cave dwelling forms (cavefish) with various degrees of reduced eyes and pigmentation. The craniofacial features we examined are the tooth-bearing maxillary bones, the nasal and antorbital bones, the circumorbital bones, and the opercular bones, all of which show evolutionary modifications in different cavefish populations. Manipulations of eye formation by transplantation of the embryonic lens, by lentectomy, or by removing the optic vesicle showed that eye-dependent and -independent processes change both the surface fish and cavefish craniofacial skeletons. The size of the olfactory pits, which the nasal and antorbital bones define, and the size and positioning of the circumorbital bones were found to correlate with eye development. For the six suborbital bones (SO1-6), the relationship with the developing eye appears to be due to ossification initiated from foci in the suborbital canal of cranial neuromasts, whose patterning is also highly correlated with the presence or absence of an eye. By contrast, we found that the number of maxillary teeth, the number of SO3 bone elements, the positioning of SO4-6 with respect to the opercular bone, and the shape of the opercular bone are not dependent on eye formation and vary among different cavefish populations. The results suggest that evolution of the cavefish craniofacial skeleton is controlled by multiple developmental events, some a direct consequence of eye degeneration and others unrelated to loss of the eye.     

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.