Use of micro-computed tomography imaging and porosity measurements as indicators of collagen preservation in archaeological bone

Collagen isolated from archaeological bone is a common material for radiocarbon dating, stable isotope analysis, and zooarchaeology by mass spectrometry (ZooMS). However, not all bones contain extant collagen, leading to unnecessary destruction of unproductive bones and wasted laboratory time and resources. An aim of this research is to study bone diagenesis, particularly collagen destruction, in an effort to develop a minimally destructive method for identifying bones with high collagen content. In a multi-method study of variably preserved bones from Etton, Cambridgeshire, UK, we examined material properties of Neolithic cattle and sheep bones including porosity, surface area, and elemental composition. Micro-computed tomography (microCT) is an imaging technique that furnishes three-dimensional images of mineralized materials such as bone. Cortical bone porosity, the percentage of total bone volume consisting of empty space as calculated using microCT, can act as a proxy for bone collagen preservation. In general, bones with high cortical porosity are unlikely to contain sufficient collagen for further analysis. Bones with apparently low cortical porosity have a more varied range of collagen preservation. Bone samples with low porosity and no extant collagen often contain micropores with a diameter of 10 nm or less that cannot be seen in microCT images but are apparent in pore size distributions measured by mercury porosimetry, and indicated by high surface areas measured by nitrogen adsorption. Furthermore, a re-evaluation of light-induced breakdown spectroscopy data from this same assemblage confirms that ratios of calcium to fluorine may likewise indicate the state of diagenesis

Is the Shroud of Turin in Relation to the Old Jerusalem Historical Earthquake?

Phillips and Hedges suggested, in the scientific magazine Nature (1989), that neutron radiation could be liable of a wrong radiocarbon dating, while proton radiation could be responsible of the Shroud body image formation. On the other hand, no plausible physical reason has been proposed so far to explain the radiation source origin, and its effects on the linen fibres. However, some recent studies, carried out by the first author and his Team at the Laboratory of Fracture Mechanics of the Politecnico di Torino, found that it is possible to generate neutron emissions from very brittle rock specimens in compression through piezonuclear fission reactions. Analogously, neutron flux increments, in correspondence to seismic activity, should be a result of the same reactions. A group of Russian scientists measured a neutron flux exceeding the background level by three orders of magnitude in correspondence to rather appreciable earthquakes (4th degree in Richter Scale). The authors consider the possibility that neutron emissions by earthquakes could have induced the image formation on Shroud linen fibres, trough thermal neutron capture by Nitrogen nuclei, and provided a wrong radiocarbon dating due to an increment in C(14,6)content. Let us consider that, although the calculated integral flux of 10^13 neutrons per square centimetre is 10 times greater than the cancer therapy dose, nevertheless it is100 times smaller than the lethal dose.Comment: 13 pages, 1 figur

Effects of spoilage on nitrogen and carbon stable isotopes signatures of the clam Ruditapes decussatus

Fish and seafood products are highly susceptible to post-mortem spoilage due to autolytic reactions at start, then microbiological activity and eventually oxidative reactions. Chemical and microbiological parameters are usually used to assess quality and make decisions for protecting public health, but they lack precision in defining which spoilage pathway is occurring at each moment. The objective of this work was to assess the effects of spoilage reactions on nitrogen and carbon stable isotopes in the grooved carpet shell clam, Ruditapes decussatus, and compare them to biochemical indicators of seafood deterioration, in order to better understand the relations between the different spoilage pathways during commercial storage conditions. Clams were kept in a refrigerator at 5 ºC, to simulate normal commercial storage conditions, and sampled in the beginning of the experiment, and after eight, ten and twelve days. Moisture, condition index, percentage edibility, total volatile basic nitrogen (TVB-N), pH, nitrogen and carbon percentages and stable isotopes were determined for each sampling moment. Stable isotope analyses were performed using a Costech Elemental Analyzer (ECS 4010) coupled to a ThermoFinnigan Delta V Advantage. Stable isotopes analysis, especially for nitrogen, proved to be a good tool for the study of clam deterioration. Nitrogen stable isotopes results showed a relation with other spoilage indicators, such as pH and TVB-N, and allowed identifying spoilage specific pathways, such as amino acids decarboxylation and production of volatile nitrogen compounds.info:eu-repo/semantics/publishedVersio

Integrating isotopes and documentary evidence : dietary patterns in a late medieval and early modern mining community, Sweden

We would like to thank the Archaeological Research Laboratory, Stockholm University, Sweden and the Tandem Laboratory (Ångström Laboratory), Uppsala University, Sweden, for undertaking the analyses of stable nitrogen and carbon isotopes in both human and animal collagen samples. Also, thanks to Elin Ahlin Sundman for providing the δ13C and δ15N values for animal references from Västerås. This research (Bäckström’s PhD employment at Lund University, Sweden) was supported by the Berit Wallenberg Foundation (BWS 2010.0176) and Jakob and Johan Söderberg’s foundation. The ‘Sala project’ (excavations and analyses) has been funded by Riksens Clenodium, Jernkontoret, Birgit and Gad Rausing’s Foundation, SAU’s Research Foundation, the Royal Physiographic Society of Lund, Berit Wallenbergs Foundation, Åke Wibergs Foundation, Lars Hiertas Memory, Helge Ax:son Johnson’s Foundation and The Royal Swedish Academy of Sciences.Peer reviewedPublisher PD

Changing environments during the Middle-Upper Palaeolithic transition in the eastern Cantabrian Region (Spain): direct evidence from stable isotope studies on ungulate bones

Environmental change has been proposed as a factor that contributed to the extinction of the Neanderthals in Europe during MIS3. Currently, the different local environmental conditions experienced at the time when Anatomically Modern Humans (AMH) met Neanderthals are not well known. In the Western Pyrenees, particularly, in the eastern end of the Cantabrian coast of the Iberian Peninsula, extensive evidence of Neanderthal and subsequent AMH activity exists, making it an ideal area in which to explore the palaeoenvironments experienced and resources exploited by both human species during the Middle to Upper Palaeolithic transition. Red deer and horse were analysed using bone collagen stable isotope analysis to reconstruct environmental conditions across the transition. A shift in the ecological niche of horses after the Mousterian demonstrates a change in environment, towards more open vegetation, linked to wider climatic change. In the Mousterian, Aurignacian and Gravettian, high inter-individual nitrogen ranges were observed in both herbivores. This could indicate that these individuals were procured from areas isotopically different in nitrogen. Differences in sulphur values between sites suggest some variability in the hunting locations exploited, reflecting the human use of different parts of the landscape. An alternative and complementary explanation proposed is that there were climatic fluctuations within the time of formation of these archaeological levels, as observed in pollen, marine and ice cores.This research was funded by the European Commission through a Marie Curie Career Integration Grant (FP7- PEOPLE-2012-CIG-322112), by the Spanish Ministry of Economy and Competitiveness (HAR2012-33956 and Ramon y Cajal-2011-00695), the University of Cantabria and Campus International to ABMA. Radiocarbon dating at ORAU was funded by MINECO-HAR2012-33956 project. J.J was supported initially by the FP7- PEOPLE-2012-CIG-322112 and later by a Marie Curie Individual Fellowship (H2020-MSCA-IF-2014-656122). Laboratory work, associated research expenses and isotopic analysis were kindly funded by the Max Planck Society to M.R

Anthropic resource exploitation and use of the territory at the onset of social complexity in the Neolithic-Chalcolithic Western Pyrenees: a multi-isotope approach

Carbon (δ13C) and nitrogen (δ15N) stable isotope analyses from bone collagen provide information about the dietary protein input, while strontium isotopes (87Sr/86Sr) from tooth enamel give us data about provenance and potential territorial mobility of past populations. To date, isotopic results on the prehistory of the Western Pyrenees are scarce. In this article, we report human and faunal values of the mentioned isotopes from the Early-Middle Neolithic site of Fuente Hoz (Anuntzeta) and the Late Neolithic/Early Chalcolithic site of Kurtzebide (Letona, Zigoitia). The main objectives of this work are to analyze the dietary and territorial mobility patterns of these populations. Furthermore, as an additional aim, we will try to discuss social ranking based on the isotope data and existing literature on this topic in the region of study. Our results show that, based on the bioavailable Sr values, both purported local and non-local humans were buried together at the sites. Additionally, they suggest similar resource consumption based on C3 terrestrial resources (i.e. ovicaprids, bovids, and suids) as the main part of the protein input. Overall, this study sheds light on how individuals from different backgrounds were still buried together and shared the same dietary lifestyle at a time in the Prehistory of Iberia when social complexities started to appear

Comparative analysis of bones, mites, soil chemistry, nematodes and soil micro-Eukaryotes from a suspected homicide to estimate the post-mortem interval

Criminal investigations of suspected murder cases require estimating the post-mortem interval (PMI, or time after death) which is challenging for longer periods. Here we present the case of human remains found in a Swiss forest. We have used a multidisciplinary approach involving the analysis of bones, soil chemical characteristics, mites and nematodes (by microscopy) and micro-Eukaryotes (by Illumina high throughput sequencing). We analysed soil samples collected beneath the remains of the head, upper and lower body and “control” samples taken a few meters away. The PMI estimated on hair 14C-data via bomb peak radiocarbon dating gave a time range of 1 to 2 years before the finding of the remains on site. Cluster analyses for chemical constituents, nematodes, mites and micro- Eukaryotes revealed two clusters 1) head and upper body and 2) lower body and controls. From mite evidence, we conclude that the body was likely to have been brought to the site after death. However, chemical analyses, nematode community analyses and the analyses of micro-Eukaryotes indicate that decomposition took place at least partly on site. This study illustrates the usefulness of combining several lines of evidence for the study of homicide cases to better calibrate PMI inference tools

Empirical Evaluation of Bone Extraction Protocols

The application of high-resolution analytical techniques to characterize ancient bone proteins requires clean, efficient extraction to obtain high quality data. Here, we evaluated many different protocols from the literature on ostrich cortical bone and moa cortical bone to evaluate their yield and relative purity using the identification of antibody-antigen complexes on enzyme-linked immunosorbent assay and gel electrophoresis. Moa bone provided an ancient comparison for the effectiveness of bone extraction protocols tested on ostrich bone. For the immunological part of this study, we focused on collagen I, osteocalcin, and hemoglobin because collagen and osteocalcin are the most abundant proteins in the mineralized extracellular matrix and hemoglobin is common in the vasculature. Most of these procedures demineralize the bone first, and then the remaining organics are chemically extracted. We found that the use of hydrochloric acid, rather than ethylenediaminetetraacetic acid, for demineralization resulted in the cleanest extractions because the acid was easily removed. In contrast, the use of ethylenediaminetetraacetic acid resulted in smearing upon electrophoretic separation, possibly indicating these samples were not as pure. The denaturing agents sodium dodecyl sulfate, urea, and guanidine HCl have been used extensively for the solubilization of proteins in non-biomineralized tissue, but only the latter has been used on bone. We show that all three denaturing agents are effective for extracting bone proteins. One additional method tested uses ammonium bicarbonate as a solubilizing buffer that is more appropriate for post-extraction analyses (e.g., proteomics) by removing the need for desalting. We found that both guanidine HCl and ammonium bicarbonate were effective for extracting many bone proteins, resulting in similar electrophoretic patterns. With the increasing use of proteomics, a new generation of scientists are now interested in the study of proteins from not only extant bone but also from ancient bone