Vittrup Man-The life-history of a genetic foreigner in Neolithic Denmark.

The lethally maltreated body of Vittrup Man was deposited in a Danish bog, probably as part of a ritualised sacrifice. It happened between c. 3300 and 3100 cal years BC, i.e., during the period of the local farming-based Funnel Beaker Culture. In terms of skull morphological features, he differs from the majority of the contemporaneous farmers found in Denmark, and associates with hunter-gatherers, who inhabited Scandinavia during the previous millennia. His skeletal remains were selected for transdisciplinary analysis to reveal his life-history in terms of a population historical perspective. We report the combined results of an integrated set of genetic, isotopic, physical anthropological and archaeological analytical approaches. Strontium signature suggests a foreign birthplace that could be in Norway or Sweden. In addition, enamel oxygen isotope values indicate that as a child he lived in a colder climate, i.e., to the north of the regions inhabited by farmers. Genomic data in fact demonstrates that he is closely related to Mesolithic humans known from Norway and Sweden. Moreover, dietary stable isotope analyses on enamel and bone collagen demonstrate a fisher-hunter way of life in his childhood and a diet typical of farmers later on. Such a variable life-history is also reflected by proteomic analysis of hardened organic deposits on his teeth, indicating the consumption of forager food (seal, whale and marine fish) as well as farmer food (sheep/goat). From a dietary isotopic transect of one of his teeth it is shown that his transfer between societies of foragers and farmers took place near to the end of his teenage years.

Elevated genetic risk for multiple sclerosis emerged in steppe pastoralist populations.

Multiple sclerosis (MS) is a neuro-inflammatory and neurodegenerative disease that is most prevalent in Northern Europe. Although it is known that inherited risk for MS is located within or in close proximity to immune-related genes, it is unknown when, where and how this genetic risk originated1. Here, by using a large ancient genome dataset from the Mesolithic period to the Bronze Age2, along with new Medieval and post-Medieval genomes, we show that the genetic risk for MS rose among pastoralists from the Pontic steppe and was brought into Europe by the Yamnaya-related migration approximately 5,000 years ago. We further show that these MS-associated immunogenetic variants underwent positive selection both within the steppe population and later in Europe, probably driven by pathogenic challenges coinciding with changes in diet, lifestyle and population density. This study highlights the critical importance of the Neolithic period and Bronze Age as determinants of modern immune responses and their subsequent effect on the risk of developing MS in a changing environment.

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.

Using SXRF and LA-ICP-TOFMS to Explore Evidence of Treatment and Physiological Responses to Leprosy in Medieval Denmark.

Leprosy can lead to blood depletion in Zn, Ca, Mg, and Fe and blood enrichment in Cu. In late medieval Europe, minerals were used to treat leprosy. Here, physiological responses to leprosy and possible evidence of treatment are investigated in enamel, dentine, and cementum of leprosy sufferers from medieval Denmark (n = 12) and early 20th century Romania (n = 2). Using SXRF and LA-ICP-TOFMS, 12 elements were mapped in 15 tooth thin sections, and the statistical covariation of paired elements was computed to assess their biological relevance. The results show marked covariations in the Zn, Ca, and Mg distributions, which are compatible with clinical studies but cannot be directly attributed to leprosy. Minerals used historically as a treatment for leprosy show no detectable intake (As, Hg) or a diffuse distribution (Pb) related to daily ingestion. Intense Pb enrichments indicate acute incorporations of Pb, potentially through the administration of Pb-enriched medication or the mobilization of Pb from bone stores to the bloodstream during intense physiological stress related to leprosy. However, comparisons with a healthy control group are needed to ascertain these interpretations. The positive correlations and the patterns observed between Pb and essential elements may indicate underlying pathophysiological conditions, demonstrating the potential of SXRF and LA-ICP-TOFMS for paleopathological investigations.

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.

How 3D printing and physical reconstruction of a skull helped in a complex pathological case.

Taphonomic changes can mimic pathological bone lesions and make it difficult to distinguish between ante- and postmortem abnormalities and to perform differential diagnostics. 3D methods, such as CT scanning and 3D printing, can be used as complementary tools to overcome the taphonomic changes. Here, we reconstructed a skull of an interesting archeological case showing severe pathological changes using 3D printing and physical reconstruction. The skeleton belonged to an approximately 18-year-old female. The bones of the entire skeleton exhibited several pathological changes. Both cranium and mandible were asymmetric, with a malformed foramen magnum and left zygomatic process, agenesis of the left external acoustic meatus, cribra orbitalia, and a bilateral absence of mental foramen. The overall diagnostic interpretation was hampered by the extensive taphonomic damage that affected the left side of the skull. In particular, the frontal bone showed taphonomic breakage and plastic deformations were present on the left side of the cranium. The skull was CT-scanned and 3D models of all the single bones were 3D printed and manually re-assembled. The intact reconstructed skull was used to estimate the endocranial volume (ECV). We showed how 3D printing and physical reconstruction of the skull helped overcome the extensive taphonomic alterations; the reconstructed skull provides important diagnostic information such as ECV and a better picture of the original ante-mortem pathological state of the skull.

Leprosy in medieval Denmark: Exploring life histories through a multi-tissue and multi-isotopic approach.

OBJECTIVES: By focusing on two Danish leprosaria (Naestved and Odense; 13th-16th c. CE) and using diet and origin as proxies, we follow a multi-isotopic approach to reconstruct life histories of patients and investigate how leprosy affected both institutionalized individuals and the medieval Danish community as a whole. MATERIALS AND METHODS: We combine archaeology, historical sources, biological anthropology, isotopic analyses (δ13 C, δ15 N, δ34 S, 87 Sr/86 Sr) and radiocarbon dating, and further analyze bones with different turnover rates (ribs and long bones). RESULTS: The δ13 C, δ15 N and δ34 S results indicate a C3 terrestrial diet with small contributions of marine protein for leprosy patients and individuals from other medieval Danish sites. A similar diet is seen through time, between males and females, and patients with and without changes on facial bones. The isotopic comparison between ribs and long bones reveals no significant dietary change. The δ34 S and 87 Sr/86 Sr results suggest that patients were local to the regions of the leprosaria. Moreover, the radiocarbon dates show a mere 50% agreement with the arm position dating method used in Denmark. CONCLUSIONS: A local origin for the leprosy patients is in line with historical evidence, unlike the small dietary contribution of marine protein. Although only 10% of the analyzed individuals have rib/long bone offsets that undoubtedly show a dietary shift, the data appear to reveal a pattern for 25 individuals (out of 50), with elevated δ13 C and/or δ15 N values in the ribs compared to the long bones, which points toward a communal type of diet and reveals organizational aspects of the institution.

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.

Mapping human mobility during the third and second millennia BC in present-day Denmark.

We present results of the largest multidisciplinary human mobility investigation to date of skeletal remains from present-day Denmark encompassing the 3rd and 2nd millennia BC. Through a multi-analytical approach based on 88 individuals from 37 different archaeological localities in which we combine strontium isotope and radiocarbon analyses together with anthropological investigations, we explore whether there are significant changes in human mobility patterns during this period. Overall, our data suggest that mobility of people seems to have been continuous throughout the 3rd and 2nd millennia BC. However, our data also indicate a clear shift in mobility patterns from around 1600 BC onwards, with a larger variation in the geographical origin of the migrants, and potentially including more distant regions. This shift occurred during a transition period at the beginning of the Nordic Bronze Age at a time when society flourished, expanded and experienced an unprecedented economic growth, suggesting that these aspects were closely related.

Wounded to the bone: Digital microscopic analysis of traumas in a medieval mass grave assemblage (Sandbjerget, Denmark, AD 1300-1350).

Battle-related mass burials are considered the most unequivocal evidence of past violence. However, most published studies involve only macroscopic analysis of skeletal remains, commonly arriving only at broad conclusions regarding trauma interpretation. The current study considers a possible avenue for achieving both greater detail and accuracy through digital microscopy. Patterns of injury were investigated among 45 individuals from a Medieval Danish mass grave (Sandbjerget, AD 1300-1350). Injuries were recorded on every anatomical element, except hand and foot bones. Each was photographed and cast, facilitating remote evaluations. Macroscopic analysis was compared with digital microscopy in order to test the relative utility of the latter in characterizing skeletal injuries (mechanism, weapon class, direction, timing of injury). The location of 201 observed injuries, mainly sharp force defects, suggested that many lesions were probably not inflicted by face-to-face opponents. Some microscopic features were indicative of a specific lesion type and weapon class. Digital microscopy was therefore demonstrated to be a complementary tool to macroscopic assessment, enhancing feature observation and quantification and serving to compensate for many of the limitations of macroscopic assessment.

A matter of months: High precision migration chronology of a Bronze Age female.

Establishing the age at which prehistoric individuals move away from their childhood residential location holds crucial information about the socio dynamics and mobility patterns in ancient societies. We present a novel combination of strontium isotope analyses performed on the over 3000 year old "Skrydstrup Woman" from Denmark, for whom we compiled a highly detailed month-scale model of her migration timeline. When combined with physical anthropological analyses this timeline can be related to the chronological age at which the residential location changed. We conducted a series of high-resolution strontium isotope analyses of hard and soft human tissues and combined these with anthropological investigations including CT-scanning and 3D visualizations. The Skrydstrup Woman lived during a pan-European period characterized by technical innovation and great social transformations stimulated by long-distance connections; consequently she represents an important part of both Danish and European prehistory. Our multidisciplinary study involves complementary biochemical, biomolecular and microscopy analyses of her scalp hair. Our results reveal that the Skrydstrup Woman was between 17-18 years old when she died, and that she moved from her place of origin -outside present day Denmark- to the Skrydstrup area in Denmark 47 to 42 months before she died. Hence, she was between 13 to 14 years old when she migrated to and resided in the area around Skrydstrup for the rest of her life. From an archaeological standpoint, this one-time and one-way movement of an elite female during the possible "age of marriageability" might suggest that she migrated with the aim of establishing an alliance between chiefdoms. Consequently, this detailed multidisciplinary investigation provides a novel tool to reconstruct high resolution chronology of individual mobility with the perspective of studying complex patterns of social and economic interaction in prehistory.

Stature in 19th and early 20th century Copenhagen. A comparative study based on skeletal remains.

Individual stature depends on multifactorial causes and is often used as a proxy for investigating the biological standard of living. While the majority of European studies on 19th and 20th century populations are based on conscript heights, stature derived from skeletal remains are scarce. For the first time in Denmark this study makes a comparison between skeletal stature and contemporary Danish conscript heights and investigates stature of males and females temporally and between socially distinct individuals and populations in 19th and early 20th century Copenhagen. A total of 357 individuals (181 males, 176 females) excavated at the Assistens cemetery in Copenhagen is analyzed. Two stature regression formulae (Trotter, 1970; Boldsen, 1990) are applied using femur measurements and evaluated compared to conscript heights. The results indicate that mean male stature using Boldsen follows a similar trend as the Danish conscript heights and that Trotter overestimate stature by ca. 6cm over Boldsen. At an inter population level statistically significant differences in male stature are observed between first and second half of the 19th century towards a slight stature decrease and larger variation while there are no significant changes observed in female stature. There are insignificant differences in stature between middle and high class individuals, but male stature differs statistically between cemeteries (p=0.000) representing middle/high class, paupers and navy employees, respectively. Female stature had no significant wealth gradient (p=0.516). This study provides new evidence of stature among males and females during the 19th century and suggests that males may have been more sensitive to changes in environmental living and nutrition than females.

Uniform diet in a diverse society. Revealing new dietary evidence of the Danish Roman Iron Age based on stable isotope analysis.

A systematic dietary investigation during Danish Roman Iron Age (1-375AD) is conducted by analyzing stable isotope ratios of carbon (δ(13) C) and nitrogen (δ(15) N) in the collagen of human and animal bone. The human sample comprises 77 individuals from 10 burial sites. In addition 31 samples of mammals and fish were analyzed from same geographical area. The investigation characterizes the human diet among different social groupings and analyses dietary differences present between sex, age, and site phase groups. Diachronically, the study investigates the Roman influences that had an effect on social structure and subsistence economy in both the Early and Late Period. Geographically the locations are both inland and coastal. The isotopic data indicate extremely uniform diet both between and within population groups from Early and Late Roman periods and the data are consistent throughout the Roman Iron Age. Protein consumption was dominated by terrestrial animals with no differences among social status, age, sex, or time period, while terrestrial plant protein only seems to have contributed little in the diet. Furthermore, the consumption of marine or aquatic resources does not seem to have been important, even among the individuals living next to the coast.

The petrous bone--a new sampling site for identifying early dietary patterns in stable isotopic studies.

Intraskeletal variation in the composition of carbon (delta(13)C) and nitrogen (delta(15)N) stable isotopes measured in collagen is tested from various human bones and dentine. Samples were taken from the femur, rib, and petrous part of the temporal bone from well-preserved skeletons of both adults (n = 34) and subadults (n = 24). Additional samples of dentine from the root of 1st molars were taken from 16 individuals. The skeletal material is from a medieval cemetery (AD 1200-1573) in Holbaek, Denmark. Our results indicate that the petrous bone has an isotopic signal that differs significantly from that of femur and rib within the single skeleton (P < 0.001 and P < 0.001, respectively), with only minor variation seen between femur and rib. On the other hand, there was no significant difference between the petrous bone and the 1st molar. The intraskeletal variation may reflect differences in turnover rate among skeletal elements. The inner periosteal layer of the petrous bone is formed in uterus and does not undergo any further remodelling after the age of 2 years, whereas the rib and femur have a continuous turnover rate of approximately 5 and 10-20 years, respectively. From the results of this study it is believed the petrous bone may be a new useful bone element and a supplement or a proxy for teeth in the analysis of early dietary patterns as it may reflect diet in fetal stages and early years of life.