Detection of chromosomal aneuploidy in ancient genomes.

Ancient DNA is a valuable tool for investigating genetic and evolutionary history that can also provide detailed profiles of the lives of ancient individuals. In this study, we develop a generalised computational approach to detect aneuploidies (atypical autosomal and sex chromosome karyotypes) in the ancient genetic record and distinguish such karyotypes from contamination. We confirm that aneuploidies can be detected even in low-coverage genomes ( ~ 0.0001-fold), common in ancient DNA. We apply this method to ancient skeletal remains from Britain to document the first instance of mosaic Turner syndrome (45,X0/46,XX) in the ancient genetic record in an Iron Age individual sequenced to average 9-fold coverage, the earliest known incidence of an individual with a 47,XYY karyotype from the Early Medieval period, as well as individuals with Klinefelter (47,XXY) and Down syndrome (47,XY, + 21). Overall, our approach provides an accessible and automated framework allowing for the detection of individuals with aneuploidies, which extends previous binary approaches. This tool can facilitate the interpretation of burial context and living conditions, as well as elucidate past perceptions of biological sex and people with diverse biological traits.

Large-scale violence in Late Neolithic Western Europe based on expanded skeletal evidence from San Juan ante Portam Latinam.

This paper explores the nature and extent of conflict in Late Neolithic Europe based on expanded skeletal evidence for violence from the San Juan ante Portam Latinam rockshelter in present-day Spain (ca. 3380-3000 cal. BC). The systematic osteological re-examination has identified 65 unhealed and 89 healed traumas-of which 77 were previously undocumented-consistent with aggression. They affect 23.1% of the 338 individuals represented. Adolescent and adult males are particularly affected (44.9% of the 107 identified), comprising 97.6% of unhealed trauma and 81.7% of healed trauma recorded in individuals whose sex could be estimated and showing higher frequencies of injuries per individual than other demographic subgroups. Results suggest that many individuals, essentially men, were exposed to violence and eventually killed in battle and raids, since warriorship is mainly restricted to this demographic in many societies. The proportion of casualties is likely to have been far greater than indicated by the 10.1% individuals exhibiting unhealed trauma, given the presence of isolated cases of unhealed postcranial trauma and of arrowheads potentially having impacted into soft tissues. This, together with skeletal indicators of poor health and the possible socioeconomic outcomes evidenced in the region, suggest wider social impacts, which may relate to a more sophisticated and formalized way of warfare than previously appreciated in the European Neolithic record.

Yersinia pestis genomes reveal plague in Britain 4000 years ago.

Extinct lineages of Yersinia pestis, the causative agent of the plague, have been identified in several individuals from Eurasia between 5000 and 2500 years before present (BP). One of these, termed the 'LNBA lineage' (Late Neolithic and Bronze Age), has been suggested to have spread into Europe with human groups expanding from the Eurasian steppe. Here, we show that the LNBA plague was spread to Europe's northwestern periphery by sequencing three Yersinia pestis genomes from Britain, all dating to ~4000 cal BP. Two individuals were from an unusual mass burial context in Charterhouse Warren, Somerset, and one individual was from a single burial under a ring cairn monument in Levens, Cumbria. To our knowledge, this represents the earliest evidence of LNBA plague in Britain documented to date. All three British Yersinia pestis genomes belong to a sublineage previously observed in Bronze Age individuals from Central Europe that had lost the putative virulence factor yapC. This sublineage is later found in Eastern Asia ~3200 cal BP. While the severity of the disease is currently unclear, the wide geographic distribution within a few centuries suggests substantial transmissibility.

Provenancing antiquarian museum collections using multi-isotope analysis.

Many of the most significant archaeological sites in Europe were excavated by antiquarians over one hundred years ago. Modern museum collections therefore frequently contain human remains that were recovered during the nineteenth and early twentieth centuries. Here we apply multi-isotope analysis (87Sr/86Sr, δ 18O, δ 13C, δ 15N) and 14C dating to evaluate the provenance of human remains within a collection that is thought to have been recovered from one of the most important archaeological sites in Britain. Excavated in 1910, the site of Coldrum in Kent is a megalithic burial monument that may be one of the earliest sites associated with the transition to farming in Britain. The interpretation of this site is therefore key to understanding how agriculture began. Using isotope analysis we show that although the human skeletal collections attributed to Coldrum do contain some of the earliest dated Neolithic human remains in Britain, they also contain the remains of individuals of fifth to seventh centuries AD date. We evaluate subsistence and mobility patterns of early Neolithic populations and provide new information about the origins of those individuals in the collection that date to the fifth to seventh centuries AD. We demonstrate the utility of employing isotope analysis to provide direct and independent information about the provenance of human remains in museum collections.

A sting in the tail: An embedded stingray spine in a mid-1st millennium AD adult male skeleton from Rebun Island, Hokkaido, Japan.

OBJECTIVES: We report here a stingray spine (Dasyatidae) found embedded in the femur of a male skeleton from the archaeological site of Uedomari-5, Rebun Island, Hokkaido, Japan. MATERIALS: A single well-preserved but incomplete human skeleton. METHODS: Macroscopic observation and low power magnification, CT imaging, radiocarbon dating and stable isotope (carbon, nitrogen) analysis. RESULTS: The stingray spine is tentatively identified as Bathytoshia brevicaudata. CT imaging shows no healing, indicating that death occurred shortly afterwards. The skeleton has been directly radiocarbon dated to the Okhotsk period (cal AD 429-827), with δ13C (-13.7‰) and δ15N (19.3‰) values indicating a diet focused on marine foods. CONCLUSIONS: The absence of healing in what would have been a non-lethal injury strongly suggests that the spine tipped an arrowhead, rather than being the result of an accidental encounter with a living stingray. It is possible that the injury reflects a period of increased conflict coinciding with, or following on from, the expansion of the Okhotsk culture from Sakhalin into northern Hokkaido. SIGNIFICANCE: Uedomari-5 provides the first example, to our knowledge, of a stingray spine directly embedded in human bone at an archaeological site. More widely, the finding contributes to our knowledge of conflict in northern hunter-gatherer communities. LIMITATIONS: Given the early excavation date (1949-50), there is little contextual information available for the burials. SUGGESTIONS FOR FURTHER RESEARCH: ZooMS (Zooarchaeology by Mass Spectrometry) may be able to identify the stingray species. Archival research may provide more information concerning the excavations at Uedomari-5.

Micromilling vs hand drilling in stable isotope analyses of incremental carbonates: The potential for δ13 C contamination by embedding resin.

RATIONALE: Embedding resins are commonly used to facilitate high-resolution sampling for stable isotope analysis but anomalous δ13 C values have been observed in some cases. Here we compare the results of microsampling strategies for hand-drilled versus resin-embedded micromilled samples from the same marine shells to assess whether resin contamination is implicated in δ13 C spikes. The comparison allows assessment of the relative benefits for spatial resolution, seasonal range for both δ18 O and δ13 C, and sample failure rates. METHODS: Hand-drilled samples were obtained from two bivalve shells (Spisula sachalinensis), corresponding to micromilled samples on the same shells where high δ13 C spikes were observed. All carbonate powders were analysed using a dual-inlet Isoprime mass spectrometer and Multiprep device. Results from both sample sets were compared statistically. RESULTS: No anomalous high δ13 C values and no failures due to insufficient gas were observed in the hand-drilled samples in contrast to the embedded micromilled sequences. Spatial resolution was reduced (~2.5×) in the former compared with the latter, resulting in a small reduction in the total range observed in the micromilled δ13 C and δ18 O values. Reduced sampling resolution between the two datasets was only significant for δ18 O. CONCLUSIONS: For S. sachalinensis (as with other similar bivalves), rapid growth mitigates the reduced sampling resolution of hand drilling and does not significantly impact observed isotopic range and seasonal patterning. Occurrence of anomalous δ13 C values were eliminated and failure rates due to insufficient sample size greatly reduced in the hand-drilled dataset. We can find no other explanation for the occurrence of δ13 C spikes than contamination by the embedding agent. We conclude that the logistical and interpretational benefits of careful hand drilling may be preferable to resin embedding for micromilling in marine shells, corals or speleothems where growth rate is rapid and the highest resolution is not required.

Physicochemical Changes in Bone Bioapatite During the Late Postmortem Interval Pre- and Post-Burning.

Postmortem chemical transformation of bone bioapatite can take place during early diagenesis, resulting in a more thermodynamically stable mineral phase. This paper examines the impact of a one year postmortem interval on unburnt and burnt bone's structural and chemical alterations. This question is of importance for the reconstruction of funerary practices involving cremation in the archaeological record, as well as forensic anthropological investigations. Fleshed pig (Sus scrofa) tibiae were left exposed in a field, then collected at 14, 34, 91, 180, and 365 day intervals prior to being burnt in an outdoor fire (≤750 °C bone temperature). Fresh (fleshed) tibiae acted as unburnt and burnt controls. Also included in the study were two cremated human bone fragments from Middle-Late Neolithic (ca. 3300-2500 BCE) Ireland. Samples were analyzed for major and trace elements using an electron microprobe wavelength dispersive analyzer and molecular structures using Fourier transform infrared spectroscopy. Linear regression, principal component analysis, linear discriminant analysis, and multivariate analysis of variance were performed for statistical analysis. Results indicate that the concentrations of elements associated with extracellular fluid (K, Na, and Cl) change with the postmortem interval (PMI) and survive burning. K values under 0.07 ± 0.01 wt% in the inner and mid-cortical zones of burnt bones suggest that bones were not burnt immediately after death. Using this criterion, results from the archaeological samples would indicate a PMI of at least weeks to months prior to cremation. Ca, P, Fe, Al, Si, and Sr are not significantly altered with burning, and Fe, Al, Si, and Sr are also unaffected by the PMI. In unburnt bones increased crystallinity and carbonate loss are detectable in <1 year, but both are obscured by burning. Structurally, the carbonate to phosphate ratio (C/P), the phosphate high temperature, and cyanamide to phosphate (CN/P) are the most useful ratios for discriminating between unburnt and burnt bones.

Radiocarbon dating from Yuzhniy Oleniy Ostrov cemetery reveals complex human responses to socio-ecological stress during the 8.2 ka cooling event.

Yuzhniy Oleniy Ostrov in Karelia, northwest Russia, is one of the largest Early Holocene cemeteries in northern Eurasia, with 177 burials recovered in excavations in the 1930s; originally, more than 400 graves may have been present. A new radiocarbon dating programme, taking into account a correction for freshwater reservoir effects, suggests that the main use of the cemetery spanned only some 100-300 years, centring on ca. 8250 to 8000 cal BP. This coincides remarkably closely with the 8.2 ka cooling event, the most dramatic climatic downturn in the Holocene in the northern hemisphere, inviting an interpretation in terms of human response to a climate-driven environmental change. Rather than suggesting a simple deterministic relationship, we draw on a body of anthropological and archaeological theory to argue that the burial of the dead at this location served to demarcate and negotiate rights of access to a favoured locality with particularly rich and resilient fish and game stocks during a period of regional resource depression. This resulted in increased social stress in human communities that exceeded and subverted the 'normal' commitment of many hunter-gatherers to egalitarianism and widespread resource sharing, and gave rise to greater mortuary complexity. However, this seems to have lasted only for the duration of the climate downturn. Our results have implications for understanding the context of the emergence-and dissolution-of socio-economic inequality and territoriality under conditions of socio-ecological stress.

'White gold' guano fertilizer drove agricultural intensification in the Atacama Desert from AD 1000.

The archaeological record shows that large pre-Inca agricultural systems supported settlements for centuries around the ravines and oases of northern Chile's hyperarid Atacama Desert. This raises questions about how such productivity was achieved and sustained, and its social implications. Using isotopic data of well-preserved ancient plant remains from Atacama sites, we show a dramatic increase in crop nitrogen isotope values (δ15N) from around AD 1000. Maize was most affected, with δ15N values as high as +30‰, and human bone collagen following a similar trend; moreover, their carbon isotope values (δ13C) indicate a considerable increase in the consumption of maize at the same time. We attribute the shift to extremely high δ15N values-the highest in the world for archaeological plants-to the use of seabird guano to fertilize crops. Guano-'white gold' as it came to be called-thus sustained agricultural intensification, supporting a substantial population in an otherwise extreme environment.

Towards a biologically available strontium isotope baseline for Ireland.

Strontium isotopes are used in archaeology, ecology, forensics, and other disciplines to study the origin of artefacts, humans, animals and food items. Strontium in animal and human tissues such as bone and teeth originates from food and drink consumed during life, leaving an isotopic signal corresponding to their geographical origin (i.e. where the plants grew, the animals grazed and the drinking water passed through). To contextualise the measurements obtained directly on animal and human remains, it is necessary to have a sound baseline of the isotopic variation of biologically available strontium in the landscape. In general, plants represent the main source of strontium for humans and animals as they usually contain much higher strontium concentrations than animal products (meat and milk) or drinking water. The observed difference between the strontium isotope composition of geological bedrock, soils and plants from the same locality warrants direct measurement of plants to create a reliable baseline. Here we present the first baseline of the biologically available strontium isotope composition for the island of Ireland based on 228 measurements on plants from 140 distinct locations. The isoscape shows significant variation in strontium isotope composition between different areas of Ireland with values as low as 0.7067 for the basalt outcrops in County Antrim and values of up to 0.7164 in the Mourne Mountains. This variability confirms the potential for studying mobility and landscape use of past human and animal populations in Ireland. Furthermore, in some cases, large differences were observed between different types of plants from the same location, highlighting the need to measure more than one plant sample per location for the creation of BASr baselines.

Author Correction: Ancient genomes indicate population replacement in Early Neolithic Britain.

In the version of this Article originally published, there were errors in the colour ordering of the legend in Fig. 5b, and in the positions of the target and surrogate populations in Fig. 5c. This has now been corrected. The conclusions of the study are in no way affected. The errors have been corrected in the HTML and PDF versions of the article.

Ancient genomes indicate population replacement in Early Neolithic Britain.

The roles of migration, admixture and acculturation in the European transition to farming have been debated for over 100 years. Genome-wide ancient DNA studies indicate predominantly Aegean ancestry for continental Neolithic farmers, but also variable admixture with local Mesolithic hunter-gatherers. Neolithic cultures first appear in Britain circa 4000 BC, a millennium after they appeared in adjacent areas of continental Europe. The pattern and process of this delayed British Neolithic transition remain unclear. We assembled genome-wide data from 6 Mesolithic and 67 Neolithic individuals found in Britain, dating 8500-2500 BC. Our analyses reveal persistent genetic affinities between Mesolithic British and Western European hunter-gatherers. We find overwhelming support for agriculture being introduced to Britain by incoming continental farmers, with small, geographically structured levels of hunter-gatherer ancestry. Unlike other European Neolithic populations, we detect no resurgence of hunter-gatherer ancestry at any time during the Neolithic in Britain. Genetic affinities with Iberian Neolithic individuals indicate that British Neolithic people were mostly descended from Aegean farmers who followed the Mediterranean route of dispersal. We also infer considerable variation in pigmentation levels in Europe by circa 6000 BC.

Strontium isotope analysis on cremated human remains from Stonehenge support links with west Wales.

Cremated human remains from Stonehenge provide direct evidence on the life of those few select individuals buried at this iconic Neolithic monument. The practice of cremation has, however, precluded the application of strontium isotope analysis of tooth enamel as the standard chemical approach to study their origin. New developments in strontium isotopic analysis of cremated bone reveal that at least 10 of the 25 cremated individuals analysed did not spend their lives on the Wessex chalk on which the monument is found. Combined with the archaeological evidence, we suggest that their most plausible origin lies in west Wales, the source of the bluestones erected in the early stage of the monument's construction. These results emphasise the importance of inter-regional connections involving the movement of both materials and people in the construction and use of Stonehenge.

Anthropogenic changes to the Holocene nitrogen cycle in Ireland.

Humans have always affected their ecosystems, but finding evidence for significant and lasting changes to preindustrial landscapes is rare. We report on human-caused changes to the nitrogen cycle in Ireland in the Bronze Age, associated with intensification of agriculture and animal husbandry that resulted in long-term changes to the nitrogen isotope values of animals (wild and domesticates) during the Holocene. Major changes to inputs and cycling of soil nitrogen occurred through deforestation, land clearance and management, and more intensive animal husbandry and cereal crop cultivation in the later Bronze Age; after this time, the Irish landscape took on its current form. Within the debate concerning the onset of the Anthropocene, our data suggest that human activity in Ireland was significant enough in the Bronze Age to have long-term impact, thereby marking a profound shift in the relationship between humans and their environment.

New radiocarbon dating and demographic insights into San Juan ante Portam Latinam, a possible Late Neolithic war grave in North-Central Iberia.

OBJECTIVES: San Juan ante Portam Latinam is one of a small number of European Neolithic sites meeting many of the archaeological criteria expected for a mass grave, and furthermore presents evidence for violent conflict. This study aims to differentiate between what is potentially a single episode of deposition, versus deposition over some centuries, or, alternatively, that resulting from a combination of catastrophic and attritional mortality. The criteria developed are intended to have wider applicability to other such proposed events. MATERIAL AND METHODS: Ten new AMS 14 C determinations on human bone from the site, together with previously available dates, are analyzed through Bayesian modeling to refine the site's chronology. This is used together with the population's demographic profile as the basis for agent-based demographic modeling. RESULTS: The new radiocarbon results, while improving the site's chronology, fail to resolve the question whether the burial represents a single event, or deposition over decades or centuries-primarily because the dates fall within the late fourth millennium BC plateau in the calibration curve. The demographic modeling indicates that the population's age and sex distribution fits neither a single catastrophic event nor a fully attritional mortality profile, but instead may partake of elements of both. DISCUSSION: It is proposed that San Juan ante Portam Latinam was used as burial place for the mainly adolescent and adult male dead of a particular or multiple violent engagements (e.g., battles), while previously or subsequently seeing use for attritional burial by other members of one or more surrounding communities dead over the course of a few generations. The overall bias towards males, particularly to the extent that many may represent conflict mortality, has implications for the structure of the surviving community, the members of which may have experienced increased vulnerability in the face of neighboring aggressors.

Living different lives: Early social differentiation identified through linking mortuary and isotopic variability in Late Neolithic/ Early Chalcolithic north-central Spain.

Variation in burial location and treatment is often observed in the prehistoric archaeological record, but its interpretation is usually highly ambiguous. Biomolecular approaches provide the means of addressing this variability in a way not previously possible, linking the lives of individuals to their funerary treatment. Here, we undertake stable carbon and nitrogen isotope analyses on a substantial sample of 166 individuals from a series of broadly contemporary Late Neolithic/ Early Chalcolithic (3500 to 2900 cal BC) mortuary monuments (El Sotillo, Alto de la Huesera, Chabola de la Hechicera and Longar) and caves (Las Yurdinas II, Los Husos I and Peña Larga) within a very spatially restricted area of north-central Spain, with sites separated by no more than 10 km on average. This spatial and temporal proximity allows us to focus on the question at the appropriate scale of analysis, avoiding confounding variables such as environmental change, diachronic trends in the subsistence economy, etc. The results demonstrate a statistically significant difference in human δ13C values between those interred in caves and those placed in monuments. The difference appears to be correlated with fine-grained environmental factors (elevation/ temperature/ precipitation), suggesting that use of the landscape was being divided at a very local scale. The reasons for this partitioning may involve differential social status (e.g. those interred in caves may be of lower standing with more restricted access to the valley's arable resources) or economic specialization (e.g. upland herding vs. valley farming) within the same community or, alternatively, different populations performing different funerary practices and following distinct subsistence economies in some respect. Our results contribute to a better understanding of the development of social differentiation and community specialisation on the scale of the immediate lived landscape.

Mobility during the neolithic and bronze age in northern ireland explored using strontium isotope analysis of cremated human bone.

OBJECTIVES: As many individuals were cremated in Neolithic and Bronze Age Ireland, they have not featured in investigations of individual mobility using strontium isotope analysis. Here, we build on recent experiments demonstrating excellent preservation of biogenic (87) Sr/(86) Sr in calcined bone to explore mobility in prehistoric Northern Ireland. MATERIALS AND METHODS: A novel method of strontium isotope analysis is applied to calcined bone alongside measurements on tooth enamel to human remains from five Neolithic and Bronze Age sites in Northern Ireland. We systematically sampled modern vegetation around each site to characterize biologically available strontium, and from this calculated expected values for humans consuming foods taken from within 1, 5, 10 and 20 Km catchments. This provides a more nuanced way of assessing human use of the landscape and mobility than the 'local' vs. 'non-local' dichotomy that is often employed. RESULTS: The results of this study 1) provide further support for the reliability of strontium isotope analysis on calcined bone, and 2) demonstrate that it is possible to identify isotopic differences between individuals buried at the same site, with some consuming food grown locally (within 1-5 Km) while others clearly consumed food from up to 50 Km away from their burial place. DISCUSSION: Hints of patterning emerge in spite of small sample numbers. At Ballynahatty, for instance, those represented by unburnt remains appear to have consumed food growing locally, while those represented by cremated remains did not. Furthermore, it appears that some individuals from Ballynahatty, Annaghmare and Clontygora either moved in the last few years of their life or their cremated remains were brought to the site. These results offer new insights into the choice behind coterminous cremation and inhumation rites in the Neolithic. Am J Phys Anthropol 160:397-413, 2016. © 2016 Wiley Periodicals, Inc.

Calcined bone provides a reliable substrate for strontium isotope ratios as shown by an enrichment experiment.

RATIONALE: Strontium isotopes ((87) Sr/(86) Sr) are used in archaeological and forensic science as markers of residence or mobility because they reflect the local geological substrate. Currently, tooth enamel is considered to be the most reliable tissue, but it rarely survives heating so that in cremations only calcined bone fragments survive. We set out to test the proposition that calcined bone might prove resistant to diagenesis, given its relatively high crystallinity, as the ability to measure in vivo (87) Sr/(86) Sr from calcined bone would greatly extend application to places and periods in which cremation was the dominant mortuary practice, or where unburned bone and enamel do not survive. METHODS: Tooth enamel and calcined bone samples were exposed to a (87) Sr-spiked solution for up to 1 year. Samples were removed after various intervals, and attempts were made to remove the contamination using acetic acid washes and ultrasonication. (87) Sr/(86) Sr was measured before and after pre-treatment on a Nu Plasma multi-collector induced coupled plasma mass spectrometer using NBS987 as a standard. RESULTS: The strontium isotopic ratios of all samples immersed in the spiked solution were strongly modified showing that significant amounts of strontium had been adsorbed or incorporated. After pre-treatment the enamel samples still contained significant amounts of (87) Sr-enriched contamination while the calcined bone fragments did not. CONCLUSIONS: The results of the artificial enrichment experiment demonstrate that calcined bone is more resistant to post-mortem exchange than tooth enamel, and that in vivo strontium isotopic ratios are retained in calcined bone.

Isotopic evidence for divergent diets and mobility patterns in the Atacama Desert, northern Chile, during the Late Intermediate Period (AD 900-1450).

The decline of the Tiwanaku state saw the emergence of two new cultures-Pica-Tarapacá and Atacama-during the Late Intermediate Period in northern Chile. Archeological evidence suggests that both groups practised maize agriculture and pastoralism, but that their interaction zones differed significantly. Marine resources are common at Pica-Tarapacá sites, even those far from coast, while Atacama sites in the desert oases and precordilleran area seem to have directed their networks towards the highlands. Here we apply stable carbon, nitrogen, and oxygen isotope analysis on human bone and enamel to test dietary patterns and residential mobility at two sites, Pica 8 and Quitor 6, representing the Pica-Tarapacá and Atacama cultures, respectively. Our results show that diet at the two sites indeed differed: significant but variable consumption of marine resources and maize is indicated at Pica 8, despite being an inland site, while diet at Quitor 6 was based mainly on terrestrial resources. The use of seabird guano and llama dung as fertilizers and extreme aridity may have contributed to the high nitrogen isotope values observed in Pica 8 humans. The δ(18) O values in Pica 8 individuals are generally lower than for Quitor in spite of its greater distance from the Andes. All three isotopes suggest the presence of at least five nonlocals in the 30 measured at Pica 8. This evidence for human mobility is consistent with the high levels of trade and interaction observed in the archeological record, and begins to quantify the degree of movement of specific individuals.

Patterns of violence-related skull trauma in Neolithic Southern Scandinavia.

This article examines evidence for violence as reflected in skull injuries in 378 individuals from Neolithic Denmark and Sweden (3,900-1,700 BC). It is the first large-scale crossregional study of skull trauma in southern Scandinavia, documenting skeletal evidence of violence at a population level. We also investigate the widely assumed hypothesis that Neolithic violence is male-dominated and results in primarily male injuries and fatalities. Considering crude prevalence and prevalence for individual bones of the skull allows for a more comprehensive understanding of interpersonal violence in the region, which is characterized by endemic levels of mostly nonlethal violence that affected both men and women. Crude prevalence for skull trauma reaches 9.4% in the Swedish and 16.9% in the Danish sample, whereas element-based prevalence varies between 6.2% for the right frontal and 0.6% for the left maxilla, with higher figures in the Danish sample. Significantly more males are affected by healed injuries but perimortem injuries affect males and females equally. These results suggest habitual male involvement in nonfatal violence but similar risks for both sexes for sustaining fatal injuries. In the Danish sample, a bias toward front and left-side injuries and right-side injuries in females support this scenario of differential involvement in habitual interpersonal violence, suggesting gendered differences in active engagement in conflict. It highlights the importance of large-scale studies for investigating the scale and context of violence in early agricultural societies, and the existence of varied regional patterns for overall injury prevalence as well as gendered differences in violence-related injuries.