Sex estimation of teeth at different developmental stages using dimorphic enamel peptide analysis.

OBJECTIVES: This study tests, for the first time, the applicability of a new method of sex estimation utilizing enamel peptides on a sample of deciduous and permanent teeth at different stages of mineralization, from nonadults of unknown sex, including perinates. MATERIALS AND METHODS: A total of 43 teeth from 29 nonadult individuals aged from 40 gestational weeks to 19 years old were analyzed. The sample included pairs of fully mineralized and just developing teeth from the same individual. The individuals were from four archaeological sites in England: Piddington (1st-2nd centuries AD), Coach Lane, Victoria Gate, and Fewston (all 18th-19th centuries). A method that identifies sex chromosome-linked isoforms of the peptide amelogenin from human tooth enamel was applied. The method utilizes a minimally destructive acid etching procedure and subsequent nano liquid chromatography tandem mass spectrometry. RESULTS: It was possible to determine the sex of 28 of the nonadult individuals sampled (males = 20, females = 8, undetermined = 1). Only one sample failed (CL9), due to insufficient mineralization of the sampled tooth enamel. Data are available via ProteomeXchange with identifier PXD021683. DISCUSSION: Sufficient peptide material to determine sex can be recovered even from the crowns of developing perinatal teeth that are not fully mineralized. The minimally destructive and inexpensive (compared to ancient DNA) nature of this procedure has significant implications for bioarchaeological studies of infancy and childhood.

At the world's edge: Reconstructing diet and geographic origins in medieval Iceland using isotope and trace element analyses.

OBJECTIVES: A multi-isotope study was conducted on individuals buried at Skriðuklaustur monastery (AD 1493-1554) to investigate their geographic origins and dietary composition. Comparative material from individuals excavated from Skeljastaðir, an inland farm site was also analyzed. MATERIALS AND METHODS: Bone collagen was extracted from 50 humans (Skriðuklaustur and Skeljastaðir) and 25 animals (Skriðuklaustur) and analyzed for δ13 C, δ15 N, and δ34 S. Dental enamel samples from 31 individuals (Skriðuklaustur) were also analyzed for 87 Sr/86 Sr, δ18 O, δ13 C, and trace elements (Pb, Sr, Zn, Ba). RESULTS: The mean value determined from individuals from Skriðuklaustur (n = 36) was δ13 C = -18.7 ± 0.8‰, δ15 N = 12.8 ± 1.1‰, and δ34 S = 9.0 ± 1.6‰, whereas at Skeljastaðir (n = 14), it was δ13 C = -20.5 ± 0.8‰, δ15 N = 7.8 ± 0.9‰, and δ34 S = 9.4 ± 1.6‰. At Skriðuklaustur, human dental enamel samples (n = 31) provided a 87 Sr/86 Sr range of 0.7060-0.7088, δ18 Ophosphate from 13.9 to 16.1‰ and δ13 Ccarbonate from -16.6 to -12.9‰. Inferred drinking water (δ18 Odw ) values range from -12.3 to -8.9‰. Sr concentrations range from 25.8 to 156.7 ppm, Ba from 0.11 to 0.81 ppm, Zn from 43.8 to 145.8 ppm, and Pb from 0.13 to 9.40 ppm. DISCUSSION: A combination of results indicates that the people from Skriðuklaustur were born in Iceland, but some lived inland during childhood while others lived closer to the coast. Since Skriðuklaustur was a hospital, these individuals may have sought medical treatment at the monastery. The δ13 C and δ15 N values determined from bone collagen indicate that the people residing at Skriðuklaustur consumed a diet high in marine protein, while those residing at Skeljastaðir exhibit values more consistent with terrestrial resources.

Sex determination of human remains from peptides in tooth enamel.

The assignment of biological sex to archaeological human skeletons is a fundamental requirement for the reconstruction of the human past. It is conventionally and routinely performed on adults using metric analysis and morphological traits arising from postpubertal sexual dimorphism. A maximum accuracy of ∼95% is possible if both the cranium and os coxae are present and intact, but this is seldom achievable for all skeletons. Furthermore, for infants and juveniles, there are no reliable morphological methods for sex determination without resorting to DNA analysis, which requires good DNA survival and is time-consuming. Consequently, sex determination of juvenile remains is rarely undertaken, and a dependable and expedient method that can correctly assign biological sex to human remains of any age is highly desirable. Here we present a method for sex determination of human remains by means of a minimally destructive surface acid etching of tooth enamel and subsequent identification of sex chromosome-linked isoforms of amelogenin, an enamel-forming protein, by nanoflow liquid chromatography mass spectrometry. Tooth enamel is the hardest tissue in the human body and survives burial exceptionally well, even when the rest of the skeleton or DNA in the organic fraction has decayed. Our method can reliably determine the biological sex of humans of any age using a body tissue that is difficult to cross-contaminate and is most likely to survive. The application of this method will make sex determination of adults and, for the first time, juveniles a reliable and routine activity in future bioarcheological and medico-legal science contexts.

Comparing apples and oranges: Why infant bone collagen may not reflect dietary intake in the same way as dentine collagen.

OBJECTIVES: Recent developments in incremental dentine analysis allowing increased temporal resolution for tissues formed during the first 1,000 days of life have cast doubt on the veracity of weaning studies using bone collagen carbon (δ13 C) and nitrogen (δ15 N) isotope ratio data from infants. Here, we compare published bone data from the well-preserved Anglo-Saxon site of Raunds Furnells, England, with co-forming dentine from the same individuals, and investigate the relationship of these with juvenile stature. The high-resolution isotope data recorded in dentine allow us to investigate the relationship of diet with juvenile stature during this critical period of life. MATERIALS AND METHODS: We compare incremental dentine collagen δ13 C and δ15 N data to published bone collagen data for 18 juveniles and 5 female adults from Anglo Saxon Raunds Furnells alongside new data for juvenile skeletal and dental age. An improvement in the method by sampling the first 0.5 mm of the sub-cuspal or sub-incisal dentine allows the isotopic measurement of dentine formed in utero. RESULTS AND DISCUSSION: δ13 C profiles for both dentine and bone are similar and more robust than δ15 N for estimating the age at which weaning foods are introduced. Our results suggest δ15 N values from dentine can be used to evaluate the maternal/in utero diet and physiology during pregnancy, and that infant dentine profiles may reflect diet PLUS an element of physiological stress. In particular, bone collagen fails to record the same range of δ15 N as co-forming dentine, especially where growth is stunted, suggesting that infant bone collagen is unreliable for weaning studies.

Land use and mobility during the Neolithic in Wales explored using isotope analysis of tooth enamel.

OBJECTIVES: The nature of land use and mobility during the transition to agriculture has often been debated. Here, we use isotope analysis of tooth enamel from human populations buried in two different Neolithic burial monuments, Penywyrlod and Ty Isaf, in south-east Wales, to examine patterns of land use and to evaluate where individuals obtained their childhood diet. MATERIALS AND METHODS: We employ strontium (87 Sr/86 Sr) and oxygen (δ18 O) and carbon (δ13 C) isotope analysis of enamel from adjacent molars. We compare strontium isotope values measured in enamel to locally bioavailable 87 Sr/86 Sr values. We combine discussion of these results with evaluation of new radiocarbon dates obtained from both sites. RESULTS: The majority of enamel samples from Penywyrlod have strontium isotope ratios above 0.7140. In contrast, the majority of those from Ty Isaf have 87 Sr/86 Sr values below 0.7140. At Penywyrlod oxygen isotope ratios range between 25.9 and 28.2 ‰ (mean 26.7 ± 0.6 ‰, 1σ, n = 15) and enamel δ13 Ccarbonate values range between -18.0 and -15.0 ‰ (mean -16.0 ± 0.8 ‰, 1σ, n = 15). At Ty Isaf oxygen isotope ratios exhibited by Neolithic individuals range between 25.4 and 27.7 ‰ (mean 26.7 ± 0.6 ‰, 1σ, n = 15) and enamel δ13 Ccarbonate values range between -16.9 and -14.9 ‰ (mean -16.0 ± 0.6 ‰, 1σ, n = 15). DISCUSSION: The strontium isotope results suggest that the majority of individuals buried at Penywyrlod did not source their childhood diet locally. One individual in this group has strontium isotope ratios that exceed all current known biosphere values within England and Wales. This individual is radiocarbon dated to the first few centuries of the 4th millennium BC, consistent with the period in which agriculture was initiated in Wales: the results therefore provide evidence for migration during the transition to farming in Wales. In contrast, all individuals sampled from Ty Isaf post-date the period in which agriculture is considered to have been initiated and could have sourced their childhood diet from the local region in which they were buried.

Infant mortality and isotopic complexity: New approaches to stress, maternal health, and weaning.

OBJECTIVES: Studies of the carbon and nitrogen stable isotope ratios (δ(13) C and δ(15) N) of modern tissues with a fast turnover, such as hair and fingernails, have established the relationship between these values in mothers and their infants during breastfeeding and weaning. Using collagen from high-resolution dentine sections of teeth, which form in the perinatal period we investigate the relationship between diet and physiology in this pivotal stage of life. MATERIALS AND METHODS: Childhood dentine collagen δ(13) C and δ(15) N profiles were produced from horizontal sections of permanent and deciduous teeth following the direction of development. These were from two 19th-century sites (n = 24) and a small number (n = 5) of prehistoric samples from Great Britain and Ireland. RESULTS: These high-resolution data exhibit marked differences between those who survived childhood and those who did not, the former varying little and the latter fluctuating widely. DISCUSSION: Breastfeeding and weaning behavior have a significant impact on the morbidity and mortality of infants and the adults they become. In the absence of documentary evidence, archaeological studies of bone collagen of adults and juveniles have been used to infer the prevalence and duration of breastfeeding. These interpretations rely on certain assumptions about the relationship between isotope ratios in the bone collagen of the adult females and the infants who have died. The data from this study suggest a more complex situation than previously proposed and the potential for a new approach to the study of maternal and infant health in past populations.

Oral histories: a simple method of assigning chronological age to isotopic values from human dentine collagen.

BACKGROUND: Stable isotope ratios of carbon (δ(13)C) and nitrogen (δ(15)N) in bone and dentine collagen have been used for over 30 years to estimate palaeodiet, subsistence strategy, breastfeeding duration and migration within burial populations. Recent developments in dentine microsampling allow improved temporal resolution for dietary patterns. AIM: A simple method is proposed which could be applied to human teeth to estimate chronological age represented by dentine microsamples in the direction of tooth growth, allowing comparison of dietary patterns between individuals and populations. The method is tested using profiles from permanent and deciduous teeth of two individuals. SUBJECTS AND METHODS: Using a diagrammatic representation of dentine development by approximate age for each human tooth (based on the Queen Mary University of London Atlas), this study estimated the age represented by each dentine section. Two case studies are shown: comparison of M1 and M2 from a 19th century individual from London, England, and identification of an unknown tooth from an Iron Age female adult from Scotland. RESULTS AND CONCLUSIONS: The isotopic profiles demonstrate that variations in consecutively-forming teeth can be aligned using this method to extend the dietary history of an individual or identify an unknown tooth by matching the profiles.

An individual with Sarmatian-related ancestry in Roman Britain.

In the second century CE the Roman Empire had increasing contact with Sarmatians, nomadic Iranian speakers occupying an area stretching from the Pontic-Caspian steppe to the Carpathian mountains, both in the Caucasus and in the Danubian borders of the empire.1,2,3 In 175 CE, following their defeat in the Marcomannic Wars, emperor Marcus Aurelius drafted Sarmatian cavalry into Roman legions and deployed 5,500 Sarmatian soldiers to Britain, as recorded by contemporary historian Cassius Dio.4,5 Little is known about where the Sarmatian cavalry were stationed, and no individuals connected with this historically attested event have been identified to date, leaving its impact on Britain largely unknown. Here we document Caucasus- and Sarmatian-related ancestry in the whole genome of a Roman-period individual (126-228 calibrated [cal.] CE)-an outlier without traceable ancestry related to local populations in Britain-recovered from a farmstead site in present-day Cambridgeshire, UK. Stable isotopes support a life history of mobility during childhood. Although several scenarios are possible, the historical deployment of Sarmatians to Britain provides a parsimonious explanation for this individual's extraordinary life history. Regardless of the factors behind his migrations, these results highlight how long-range mobility facilitated by the Roman Empire impacted provincial locations outside of urban centers.

Proteomics analysis of ancient food vessel stitching reveals >4000-year-old milk protein.

RATIONALE: The 19th century excavation of an exceptionally well-preserved Early Bronze Age high status log-coffin burial from northern England, dated to 2200-2020 BC, yielded a 'food residue' collected from the inside of an accompanying bark vessel. This residue contained fibrous stitching that was used to hold the bark walls of the vessel together, first described as animal sinews, although the surviving material clearly contains animal hairs. Protein sequencing by soft ionisation mass spectrometry should identify the proteins that constitute the material, as well as the animal species from which they derive. METHODS: Peptide mass fingerprinting (PMF) by MALDI-TOF-MS combined with liquid chromatography-ESI-LTQ-MS/MS was used to identify low-abundance proteins as well as the dominant proteins in the sample. RESULTS: These proteomics techniques revealed the dominant proteins as deriving from the fibrous keratins (both types 1 and 2) and collagens (types 1 and 3), specifically those indicative of a bovine source. However, several peptide sequences diagnostic of bovine α-S1-casein were also observed, indicating that traces of milk had been preserved within the >4000-year-old fibrous residue. CONCLUSIONS: The presence of this food vessel that once contained milk within a burial of high status is suggestive of the importance placed on these secondary products. It is perhaps more remarkable that this information was retrieved not only from material of such antiquity, but also from an excavation that occurred nearly 200 years ago.

Victims and survivors: stable isotopes used to identify migrants from the Great Irish Famine to 19th century London.

Historical evidence documents mass migration from Ireland to London during the period of the Great Irish Famine of 1845-52. The rural Irish were reliant on a restricted diet based on potatoes but maize, a C(4) plant, was imported from the United States of America in 1846-47 to mitigate against Famine. In London, Irish migrants joined a population with a more varied diet. To investigate and characterize their diet, carbon and nitrogen isotope ratios were obtained from bone collagen of 119 and hair keratin of six individuals from Lukin Street cemetery, Tower Hamlets (1843-54), and bone collagen of 20 individuals from the cemetery at Kilkenny Union Workhouse in Ireland (1847-51). A comparison of the results with other contemporaneous English populations suggests that Londoners may have elevated δ(15) N compared with their contemporaries in other cities. In comparison, the Irish group have lower δ(15) N. Hair analysis combined with bone collagen allows the reconstruction of perimortem dietary changes. Three children aged 5-15 years from Kilkenny have bone collagen δ(13) C values that indicate consumption of maize (C(4)). As maize was only imported into Ireland in quantity from late 1846 and 1847, these results demonstrate relatively rapid bone collagen turnover in children and highlight the importance of age-related bone turnover rates, and the impact the age of the individual can have on studies of short-term dietary change or recent migration. Stable light isotope data in this study are consistent with the epigraphic and documentary evidence for the presence of migrants within the London cemetery.