100 ancient genomes show repeated population turnovers in Neolithic Denmark.

Major migration events in Holocene Eurasia have been characterized genetically at broad regional scales1-4. However, insights into the population dynamics in the contact zones are hampered by a lack of ancient genomic data sampled at high spatiotemporal resolution5-7. Here, to address this, we analysed shotgun-sequenced genomes from 100 skeletons spanning 7,300 years of the Mesolithic period, Neolithic period and Early Bronze Age in Denmark and integrated these with proxies for diet (13C and 15N content), mobility (87Sr/86Sr ratio) and vegetation cover (pollen). We observe that Danish Mesolithic individuals of the Maglemose, Kongemose and Ertebølle cultures form a distinct genetic cluster related to other Western European hunter-gatherers. Despite shifts in material culture they displayed genetic homogeneity from around 10,500 to 5,900 calibrated years before present, when Neolithic farmers with Anatolian-derived ancestry arrived. Although the Neolithic transition was delayed by more than a millennium relative to Central Europe, it was very abrupt and resulted in a population turnover with limited genetic contribution from local hunter-gatherers. The succeeding Neolithic population, associated with the Funnel Beaker culture, persisted for only about 1,000 years before immigrants with eastern Steppe-derived ancestry arrived. This second and equally rapid population replacement gave rise to the Single Grave culture with an ancestry profile more similar to present-day Danes. In our multiproxy dataset, these major demographic events are manifested as parallel shifts in genotype, phenotype, diet and land use.

Population genomics of post-glacial western Eurasia.

Western Eurasia witnessed several large-scale human migrations during the Holocene1-5. Here, to investigate the cross-continental effects of these migrations, we shotgun-sequenced 317 genomes-mainly from the Mesolithic and Neolithic periods-from across northern and western Eurasia. These were imputed alongside published data to obtain diploid genotypes from more than 1,600 ancient humans. Our analyses revealed a 'great divide' genomic boundary extending from the Black Sea to the Baltic. Mesolithic hunter-gatherers were highly genetically differentiated east and west of this zone, and the effect of the neolithization was equally disparate. Large-scale ancestry shifts occurred in the west as farming was introduced, including near-total replacement of hunter-gatherers in many areas, whereas no substantial ancestry shifts happened east of the zone during the same period. Similarly, relatedness decreased in the west from the Neolithic transition onwards, whereas, east of the Urals, relatedness remained high until around 4,000 BP, consistent with the persistence of localized groups of hunter-gatherers. The boundary dissolved when Yamnaya-related ancestry spread across western Eurasia around 5,000 BP, resulting in a second major turnover that reached most parts of Europe within a 1,000-year span. The genetic origin and fate of the Yamnaya have remained elusive, but we show that hunter-gatherers from the Middle Don region contributed ancestry to them. Yamnaya groups later admixed with individuals associated with the Globular Amphora culture before expanding into Europe. Similar turnovers occurred in western Siberia, where we report new genomic data from a 'Neolithic steppe' cline spanning the Siberian forest steppe to Lake Baikal. These prehistoric migrations had profound and lasting effects on the genetic diversity of Eurasian populations.

The impact of age on the morphology of the 12th thoracic vertebral endplates.

The current article explores the aging effects on the overall morphology of the endplates of the 12th thoracic vertebra (T12), while screening for sex differences. It further evaluates the suitability of T12 for estimating age-at-death in bioarcheaological contexts. We captured the morphology of the vertebral endplates, including the formation of osteophytes, in a novel continuous quantitative manner using digital photography. 168 Greek adults from the Athens Collection were used for modeling the aging effects and another 107 individuals from two Danish archaeological assemblages for evaluation. Regression analysis is based on generalized additive models for correlating age-at-death and morphological variation. Our proposed measurement method is highly reliable (R>0.98) and the main differences observed between sexes are size related. Aging has considerable effect on the endplate morphology of the T12 with the total area of the endplate, the area of the epiphyseal rim, and the shape irregularities of the endplate's external boundary being mostly affected. Multivariate regression shows that aging effects account up to 46% of the observed variation, although with differential expression between sexes. Correct age prediction on archaeological remains reached 33% with a prominent tendency for overestimation. The morphology of the T12 endplates is influenced by age and it can provide some insight with respect to the age-at-death of unidentified individuals, especially when other skeletal age markers are unavailable. Our proposed method provides an age-estimation framework for bioarchaeological settings, especially for estimating broader age ranges, such as discriminating between young and old adults.

SPIN enables high throughput species identification of archaeological bone by proteomics.

Species determination based on genetic evidence is an indispensable tool in archaeology, forensics, ecology, and food authentication. Most available analytical approaches involve compromises with regard to the number of detectable species, high cost due to low throughput, or a labor-intensive manual process. Here, we introduce "Species by Proteome INvestigation" (SPIN), a shotgun proteomics workflow for analyzing archaeological bone capable of querying over 150 mammalian species by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Rapid peptide chromatography and data-independent acquisition (DIA) with throughput of 200 samples per day reduce expensive MS time, whereas streamlined sample preparation and automated data interpretation save labor costs. We confirm the successful classification of known reference bones, including domestic species and great apes, beyond the taxonomic resolution of the conventional peptide mass fingerprinting (PMF)-based Zooarchaeology by Mass Spectrometry (ZooMS) method. In a blinded study of degraded Iron-Age material from Scandinavia, SPIN produces reproducible results between replicates, which are consistent with morphological analysis. Finally, we demonstrate the high throughput capabilities of the method in a high-degradation context by analyzing more than two hundred Middle and Upper Palaeolithic bones from Southern European sites with late Neanderthal occupation. While this initial study is focused on modern and archaeological mammalian bone, SPIN will be open and expandable to other biological tissues and taxa.

Into the fire: Investigating the introduction of cremation to Nordic Bronze Age Denmark: A comparative study between different regions applying strontium isotope analyses and archaeological methods.

Changes in funerary practices are key to the understanding of social transformations of past societies. Over the course of the Nordic Bronze Age, funerary practices changed from inhumation to cremation. The aim of this study is to shed light on this fundamental change through a cross-examination of archaeometric provenance data and archaeological discussions of the context and layouts of early cremation graves. To this end, we conducted 19 new provenance analyses of strontium isotopes from Early Nordic Bronze age contexts in Thisted County and Zealand and Late Bronze Age contexts from Thisted County and Vesthimmerland (Denmark). These data are subsequently compared with data from other extant relevant studies, including those from Late Bronze Age Fraugde on the Danish island of Fyn. Overall, the variations within our provenience data suggest that the integration and establishment of cremation may not have had a one-to-one relationship with in-migration to Nordic Bronze Age Denmark. Moreover, there seems to be no single blanket scenario which dictated the uptake of cremation as a practice within this part of Southern Scandinavia. By addressing habitus in relation to the deposition of cremations as juxtaposed with these provenance data¸ we hypothesize several potential pathways for the uptake of cremation as a new cultural practice within the Danish Nordic Bronze Age and suggest that this may have been a highly individual process, whose tempo may have been dictated by the specificities of the region(s) concerned.

Diverse variola virus (smallpox) strains were widespread in northern Europe in the Viking Age.

Smallpox, one of the most devastating human diseases, killed between 300 million and 500 million people in the 20th century alone. We recovered viral sequences from 13 northern European individuals, including 11 dated to ~600-1050 CE, overlapping the Viking Age, and reconstructed near-complete variola virus genomes for four of them. The samples predate the earliest confirmed smallpox cases by ~1000 years, and the sequences reveal a now-extinct sister clade of the modern variola viruses that were in circulation before the eradication of smallpox. We date the most recent common ancestor of variola virus to ~1700 years ago. Distinct patterns of gene inactivation in the four near-complete sequences show that different evolutionary paths of genotypic host adaptation resulted in variola viruses that circulated widely among humans.

Stature Estimation from Postmortem CT Femoral Maximum Length in Contemporary Danish Population.

Stature estimation methods for Danish adult population have generally relied on Trotter and Gleser's and Boldsen's regression equations that are based on the skeletal remains of recent war dead American Whites, Terry Skeletal Collection, and Danish archaeological medieval skeletal materials, respectively. These equations are probably not suitable for stature estimation in contemporary Danish forensic cases. Furthermore, because postmortem computed tomography (PMCT) is now routinely performed at Danish forensic departments, equations based on PMCT, rather than measurements of defleshed bones, are needed. The aim of this study was to develop new equations for adult stature estimation based on PMCT femoral measurement. Maximum femoral length was measured on the PMCT images of 78 individuals (41 males and 37 females) aged 23-45 years. The measurement accuracy was tested on dry bones, and all the measurements were included in the inter- and intra-observer analyses. Both analyses results demonstrated the reliability of the method and data. Comparison between the living stature of the individuals and the estimates based on the equations by Trotter and Gleser and Boldsen demonstrated the unreliability of the previous equations to some extent. New regression equations were then developed and validated on a different sample of 18 Danish forensic cases. Comparisons of all the equations indicated that both the sets of previous equations underestimated the stature in the new validation dataset. The new equations developed in this study provide a reliable alternative for stature estimation in modern Danish forensic cases.