The application of amino acid racemization in the acid soluble fraction of enamel to the estimation of the age of human teeth

Estimation of age-at-death for skeletonised forensic remains is one of the most significant problems in forensic anthropology. The majority of existing morphological and histological techniques are highly inaccurate, and show a bias towards underestimating the age of older individuals. One technique which has been successful in forensic age estimation is amino acid racemization in dentine. However, this method cannot be used on remains where the post-mortem interval is greater than 20 years. An alternative approach is to measure amino acid racemization in dental enamel, which is believed to be more resistant to change post-mortem. The extent of amino acid racemization in the acid soluble fraction of the enamel proteins was determined for modem known age teeth. A strong correlation was observed between the age of the tooth and the extent of racemization. No systematic bias in the direction of age estimation errors was detected. For the majority of teeth analyzed, the presence of dental caries did not affect the results obtained. In a minority of cases, carious teeth showed a higher level of racemization than would be expected given the age of the individual. These results indicate that amino acid racemization in enamel has the potential to be used in age estimation of skeletal remains. (C) 2007 Elsevier Ireland Ltd. All rights reserved

Intra-crystalline protein diagenesis (IcPD) in Patella vulgata. Part I: Isolation and testing of the closed system

This study successfully isolates a fraction of intra-crystalline proteins from shells of the marine gastropod Patella vulgata and assesses the suitability of these proteins for IcPD (Intra-crystalline Protein Diagenesis) geochronology. We discuss the mineralogical composition of this gastropod, investigated for the first time by X-ray diffraction mapping, and use the results to inform our sampling strategy. The potential of the calcitic rim and of a bulk sample (containing both apex and rim) of the shell to act as stable repositories for the intra-crystalline proteins during diagenesis is examined. The composition and the diagenetic behaviour of the intra-crystalline proteins isolated from different locations within the shell are compared, highlighting the necessity of targeting consistent sampling positions. We induced artificial diagenesis of both intra-crystalline and whole-shell proteins by conducting high-temperature experiments in hydrous environment; this allowed us to quantify the loss of amino acids by leaching and therefore evaluate the open- or closed-system behaviour of the different fractions of proteins. The results obtained provide further confirmation that patterns of diagenesis vary according to the protein sequence, structure, and location within or outside the intra-crystalline fraction. As Patella is frequently found in the fossil record, both in archaeological and geological contexts, the application of IcPD geochronology to this biomineral opens up the possibility to obtain reliable age information from a range of sites in different areas of the world

Petrous bone diagenesis: a multi-analytical approach

© 2019 Elsevier B.V. The discovery of petrous bone as an excellent repository for ancient biomolecules has been a turning point in biomolecular archaeology, especially in aDNA research, but excessive and uncontrolled sampling could result in loss of this valuable resource for future research. This study reports on the histological (optical microscopy), physical (FTIR-ATR), elemental (CHN) and biochemical (collagen and DNA analysis) preservation of 15 human petrous bones spanning from the c. 2100 BCE to 1850 CE. Through the combined application of a number of diagenetic parameters (general histological index; infrared splitting factor; carbonate/phosphate ratio; amide/phosphate ratio; col wt%; % C, % N and C/N of whole bone and collagen; % endogenous DNA), we provide new insights into petrous bone micromorphological characteristics and diagenesis, and new evidence to enhance screening practices for aDNA and collagen analysis.MJC was supported by Danish National Research Foundation (DNRF128) and KP from the Leverhulme Trust (PLP-2012-116). MEA thanks The Danish National Resarch Foundation (DNRF94), the Lundbeck Foundation, the University of Copehagen (KU2016 programme) and the Vellux Foundations (Villum Young Investigator programme). IK would like to thank Onassis Foundation (grant no. F ZL 047-1/2015-2016), Leventis Foundation and the Greek Archaeological Committee UK (GACUK)

Dietary modulation of cortical excitation and inhibition

The balance of excitatory and inhibitory neurotransmitters in the brain affects both neural responses and behaviour in humans and animals. Here we investigated whether dietary intervention aimed at increasing levels of the inhibitory neurotransmitter GABA can influence neural responses to basic sensory stimuli. Using a steady-state EEG paradigm, we found that the neural response to visual patterns was reduced in individuals who consumed a yeast-extract product rich in substances associated with the production of GABA (glutamate and B vitamins), but not in a control group who consumed a placebo substance (N=14 per group). This demonstrates that the balance of excitation and inhibition in the brain can be influenced by dietary interventions, suggesting possible clinical benefits in conditions (e.g. epilepsy) where inhibition is abnormal

Bone diagenesis in a Mycenaean secondary burial (Kastrouli, Greece)

Abstract: This paper presents the characteristics of bone diagenesis in a secondary commingled Mycenaean burial in Kastrouli (Phocis, Greece) through the histological (light microscopy), physical (FTIR-ATR), and biochemical (collagen) analysis of seventeen human (including two petrous bones) and seven animal bones. Post-mortem modifications in bone microstructure, bioapatite, and collagen were characteristic of burial environments with seasonal groundwater and temperature fluctuations. The two human petrous bones displayed a lack of microscopic focal destruction (MFD) sites and a generally good histological preservation, but although a small sample size, did not show any better bioapatite and collagen preservation compared with human femora. Intra-site variations were defined by three main diagenetic patterns that display differences in histological modifications, crystallinity changes, and collagen degradation. These different patterns were either related to different microenvironment conditions and/or influenced by possible differences in the early taphonomic histories experienced by bones prior to secondary deposition. Further, this study highlights the importance of infrared splitting factor (IRSF), carbonate/phosphate ratio (C/P), and general histological index (GHI) for the qualitative assessment of archaeological bone, and the potential use of amide/phosphate ratio (Am/P) as a collagen predictor

Excavations at the Lower Palaeolithic Site at Elveden, Suffolk

The Lower Palaeolithic site at Elveden, Suffolk, was the subject of new excavations from 1995–1999. Excavations around the edge and in the centre of the former clay-pit revealed sediments infilling a lake basin that had formed in Lowestoft till, overlying Chalk, the till being attributed to the Anglian glaciation (MIS 12). The lake sediments contain pollen that can be assigned to pollen zones HoI and HoIIa of the early Hoxnian (MIS 11). Overlying grey clays contain ostracods, molluscs, vertebrates, and carbonate concretions. Together they are indicative of a fluvial environment in a temperate climate. AAR ratios (amino acid racemisation) on the molluscs also suggest correlation with MIS 11. Further indications of a fluvial context are indicated by thin spreads of lag gravel along opposite sides of the clay-pit, marking the edges of a channel. The gravel forms the raw material for the human industries which consist of handaxes, flake tools, flakes, and cores. Further artefacts are found in the overlying black clay, which is interpreted as a palaeosol that formed with the silting-up of the channel. The basin was further infilled with colluvial ‘brickearths’, which also contain artefacts that are probably derived from the underlying gravel. Further evidence of soil formation was identified in the ‘brickearth’. Coversands with periglacial involutions overlie the ‘brickearth’ at the top of the sequence. These probably formed in the last cold stage, the Devensian

Biomineralisation by earthworms: an investigation into the stability and distribution of amorphous calcium carbonate

Background Many biominerals form from amorphous calcium carbonate (ACC), but this phase is highly unstable when synthesised in its pure form inorganically. Several species of earthworm secrete calcium carbonate granules which contain highly stable ACC. We analysed the milky fluid from which granules form and solid granules for amino acid (by liquid chromatography) and functional group (by Fourier transform infrared (FTIR) spectroscopy) compositions. Granule elemental composition was determined using inductively coupled plasma-optical emission spectroscopy (ICP-OES) and electron microprobe analysis (EMPA). Mass of ACC present in solid granules was quantified using FTIR and compared to granule elemental and amino acid compositions. Bulk analysis of granules was of powdered bulk material. Spatially resolved analysis was of thin sections of granules using synchrotron-based μ-FTIR and EMPA electron microprobe analysis. Results The milky fluid from which granules form is amino acid-rich (≤ 136 ± 3 nmol mg−1 (n = 3; ± std dev) per individual amino acid); the CaCO3 phase present is ACC. Even four years after production, granules contain ACC. No correlation exists between mass of ACC present and granule elemental composition. Granule amino acid concentrations correlate well with ACC content (r ≥ 0.7, p ≤ 0.05) consistent with a role for amino acids (or the proteins they make up) in ACC stabilisation. Intra-granule variation in ACC (RSD = 16%) and amino acid concentration (RSD = 22–35%) was high for granules produced by the same earthworm. Maps of ACC distribution produced using synchrotron-based μ-FTIR mapping of granule thin sections and the relative intensity of the ν2: ν4 peak ratio, cluster analysis and component regression using ACC and calcite standards showed similar spatial distributions of likely ACC-rich and calcite-rich areas. We could not identify organic peaks in the μ-FTIR spectra and thus could not determine whether ACC-rich domains also had relatively high amino acid concentrations. No correlation exists between ACC distribution and elemental concentrations determined by EMPA. Conclusions ACC present in earthworm CaCO3 granules is highly stable. Our results suggest a role for amino acids (or proteins) in this stability. We see no evidence for stabilisation of ACC by incorporation of inorganic components