Descent, marriage, and residence practices of a 3,800-year-old pastoral community in Central Eurasia.

Our understanding of prehistoric societal organization at the family level is still limited. Here, we generated genome data from 32 individuals from an approximately 3,800-y-old burial mound attributed to the Bronze Age Srubnaya-Alakul cultural tradition at the site of Nepluyevsky, located in the Southern Ural region of Central Eurasia. We found that life expectancy was generally very low, with adult males living on average 8 y longer than females. A total of 35 first-degree, 40 second-degree, and 48 third-degree biological relationships connected 23 of the studied individuals, allowing us to propose a family tree spanning three generations with six brothers at its center. The oldest of these brothers had eight children with two women and the most children overall, whereas the other relationships were monogamous. Notably, related female children above the age of five were completely absent from the site, and adult females were more genetically diverse than males. These results suggest that biological relationships between male siblings played a structural role in society and that descent group membership was based on patrilineality. Women originated from a larger mating network and moved to join the men, with whom they were buried. Finally, the oldest brother likely held a higher social position, which was expressed in terms of fertility.

Ancient mitochondrial diversity reveals population homogeneity in Neolithic Greece and identifies population dynamics along the Danubian expansion axis.

The aim of the study is to investigate mitochondrial diversity in Neolithic Greece and its relation to hunter-gatherers and farmers who populated the Danubian Neolithic expansion axis. We sequenced 42 mitochondrial palaeogenomes from Greece and analysed them together with European set of 328 mtDNA sequences dating from the Early to the Final Neolithic and 319 modern sequences. To test for population continuity through time in Greece, we use an original structured population continuity test that simulates DNA from different periods by explicitly considering the spatial and temporal dynamics of populations. We explore specific scenarios of the mode and tempo of the European Neolithic expansion along the Danubian axis applying spatially explicit simulations coupled with Approximate Bayesian Computation. We observe a striking genetic homogeneity for the maternal line throughout the Neolithic in Greece whereas population continuity is rejected between the Neolithic and present-day Greeks. Along the Danubian expansion axis, our best-fitting scenario supports a substantial decrease in mobility and an increasing local hunter-gatherer contribution to the gene-pool of farmers following the initial rapid Neolithic expansion. Οur original simulation approach models key demographic parameters rather than inferring them from fragmentary data leading to a better understanding of this important process in European prehistory.

The genomic origins of the world's first farmers.

The precise genetic origins of the first Neolithic farming populations in Europe and Southwest Asia, as well as the processes and the timing of their differentiation, remain largely unknown. Demogenomic modeling of high-quality ancient genomes reveals that the early farmers of Anatolia and Europe emerged from a multiphase mixing of a Southwest Asian population with a strongly bottlenecked western hunter-gatherer population after the last glacial maximum. Moreover, the ancestors of the first farmers of Europe and Anatolia went through a period of extreme genetic drift during their westward range expansion, contributing highly to their genetic distinctiveness. This modeling elucidates the demographic processes at the root of the Neolithic transition and leads to a spatial interpretation of the population history of Southwest Asia and Europe during the late Pleistocene and early Holocene.

Trepanations in the ancient Greek colony of Akanthos: Skull surgery in the light of Hippocratic medicine.

OBJECTIVE: The study presents seven new cases of trepanations on four individuals from the ancient Greek colony of Akanthos (5th - 1st c. BC) and juxtaposes the paleopathological observations with the Hippocratic treatises. MATERIALS: A meta-analysis of 42 published trepanations on 27 individuals from Greece. METHODS: Trepanations from Akanthos were observed macroscopically, stereoscopically and by computer tomography. The meta-analysis considers the demographic, geographical and temporal distribution, the techniques, associated pathology, and survival rate. RESULTS: Trepanations were observed on four females in Akanthos. Three trepanations were performed with scraping and four with drilling. Two individuals exhibited significant signs of healing, and two survived shortly after the operation. In two cases, trepanation was practiced for treating cranial injuries. In Greek antiquity trepanation was performed predominantly in males, principally as a surgical treatment of cranial injuries. The survival rate was 62.9 %. Scraping was the oldest, most frequent and successful technique. CONCLUSIONS: Trepanation performed systematically in Greece since the Bronze Age (ca. 2,000 BC). Hippocrates who rationally conceived medicine, codified the pre-existing empirical knowledge. The trepanations from Akanthos show technical similarities but also discrepancies from the Hippocratic recommendations, confirming the mental and technical readiness of the ancient surgeon. SIGNIFICANCE: The synchronous to Hippocrates trepanations from Akanthos, provide the opportunity to compare physical evidence with the written sources. LIMITATIONS: The frequency of trepanations is affected by the representativeness and the state of the skeletal preservation. SUGGESTIONS FOR FUTURE RESEARCH: Trepanations published in previous decades, need to be reassessed with new imaging modalities. A standard recording methodology is fundamental for data comparison.

The genomic history of the Aegean palatial civilizations.

The Cycladic, the Minoan, and the Helladic (Mycenaean) cultures define the Bronze Age (BA) of Greece. Urbanism, complex social structures, craft and agricultural specialization, and the earliest forms of writing characterize this iconic period. We sequenced six Early to Middle BA whole genomes, along with 11 mitochondrial genomes, sampled from the three BA cultures of the Aegean Sea. The Early BA (EBA) genomes are homogeneous and derive most of their ancestry from Neolithic Aegeans, contrary to earlier hypotheses that the Neolithic-EBA cultural transition was due to massive population turnover. EBA Aegeans were shaped by relatively small-scale migration from East of the Aegean, as evidenced by the Caucasus-related ancestry also detected in Anatolians. In contrast, Middle BA (MBA) individuals of northern Greece differ from EBA populations in showing ∼50% Pontic-Caspian Steppe-related ancestry, dated at ca. 2,600-2,000 BCE. Such gene flow events during the MBA contributed toward shaping present-day Greek genomes.

A comparison of the anatomical and the mathematical stature estimation methods on an ancient Greek population.

Stature is a key concept in the study of human past since along with the biological information it provides overall trends about the standards of living. However, stature data on historic and prehistoric populations are still limited, especially for key temporospatial settings of antiquity such as ancient Greece. We collected osteometric data from 35 male and 33 female individuals (n = 68) from a Roman Period necropolis (146 BC-330 AD) in northern Greece and we applied the anatomical stature estimation method (Raxter et al. 2006). We compared this estimation with the results of 20 regression equations (both ordinary least squares and reduced major axis methods) and we examined the factors that affect their accuracy such as body proportions and the chronology and geography of reference series. For our analyses, we calculated the percent prediction error (%PE) produced by each regression equation for males and females separately. We introduced the total %PE to evaluate each equation's effectiveness on both sexes simultaneously. We calculated long bone ratios and the Euclidean distance between the Greek dataset and the reference series. According to the anatomical method, males from Northern Greece had a mean stature value of 168.2 cm ± 5.38 and females of 156.9 cm ± 5.27. The regression equations of Vercellotti et al. (2009) provided the best estimations in both sexes. Estimation errors (%PE) were not statistically different between the ordinary least squares and reduced major axis equations. The chronological affinity in body proportions between the target population and the reference series could provide significant evidence for the prediction of the optimal regression formulae. With this paper we provide the first osteometric dataset for the anatomical stature estimation method from ancient Greece and we suggest the most suitable regression equations for this key region of the ancient World.

Screening archaeological bone for palaeogenetic and palaeoproteomic studies.

The recovery and analysis of ancient DNA and protein from archaeological bone is time-consuming and expensive to carry out, while it involves the partial or complete destruction of valuable or rare specimens. The fields of palaeogenetic and palaeoproteomic research would benefit greatly from techniques that can assess the molecular quality prior to sampling. To be relevant, such screening methods should be effective, minimally-destructive, and rapid. This study reports results based on spectroscopic (Fourier-transform infrared spectroscopy in attenuated total reflectance [FTIR-ATR]; n = 266), palaeoproteomic (collagen content; n = 226), and palaeogenetic (endogenous DNA content; n = 88) techniques. We establish thresholds for three different FTIR indices, a) the infrared splitting factor [IRSF] that assesses relative changes in bioapatite crystals' size and homogeneity; b) the carbonate-to-phosphate [C/P] ratio as a relative measure of carbonate content in bioapatite crystals; and c) the amide-to-phosphate ratio [Am/P] for assessing the relative organic content preserved in bone. These thresholds are both extremely reliable and easy to apply for the successful and rapid distinction between well- and poorly-preserved specimens. This is a milestone for choosing appropriate samples prior to genomic and collagen analyses, with important implications for biomolecular archaeology and palaeontology.

"What and how should we share?" An inter-method inter-observer comparison of measurement error with landmark-based craniometric datasets.

The present study evaluates the precision and accuracy of photogrammetric 3D modeling of human crania in landmark acquisition and explores the limitations of combining datasets acquired by different observers and different measurement methods. Our working sample comprises 50 adult human crania, which were modeled with 3D photogrammetry. 3D coordinates of 56 landmarks were collected from the 3D models with Meshlab software and an existing corresponding dataset digitized with Microscribe-3DX has been utilized. Measurement error for landmark configurations and Inter Landmarks Distances (ILDs) for each type of landmarks has been assessed through least root mean squared deviation and mean absolute error respectively. Inter-observer error has been assessed on a sub-sample of 20 crania, which was also used for caliper measured ILDs. Between-methods Technical Error Measurement (TEM) based on ILDs has been calculated for evaluating the interchangeability for different datasets. Photogrammetric 3D models and Microscribe-3DX share identical rated accuracy regarding craniometric applications and both methods show increased accuracy in locating type I landmarks as opposed to types II and III. However, photogrammetric 3D models perform better in terms of inter-observer error suggesting higher reliability of measurements. Furthermore, ILDs are less prone to measurement error than landmark configurations. Finally, ILDs exhibit similar relative TEM of about 1.5% between Microscribe, caliper and 3D model based measurement methods. Combining datasets of landmark coordinates acquired from photogrammetric 3D models does not compromise the statistical integrity in terms of measurement error, which also applies to pooling ILD datasets from multiple methods. Nevertheless, compiling 3D datasets from multiple methods for 3DGM analysis should be done cautiously.

Ongoing tissue changes in an experimentally mummified human leg.

Artificial mummification has been used since antiquity and is best known from ancient Egypt. Despite ancient Egyptian mummies being studied for several decades, the mummification techniques of that time are not well understood. Modern mummification experiments involving animal and human tissues have contributed additional insights relevant to a broad field of research. In the current study, we present follow-up results of an experiment on artificial mummification, which began in 2009. A human leg was artificially mummified and monitored for almost a year with histological, molecular, and radiological techniques. Since then, it has remained in a dry, natron salt blend for 9 years. The current analyses show further progression of dehydration and tissue alterations, as well as DNA degradation, suggesting an ongoing process. Our results add new insights into the mechanisms of tissue mummification. Taking into account that the process is still ongoing, further research is required, including a re-evaluation of the human leg in the future.

Bayesian estimation of partial population continuity using ancient DNA and spatially explicit simulations.

The retrieval of ancient DNA from osteological material provides direct evidence of human genetic diversity in the past. Ancient DNA samples are often used to investigate whether there was population continuity in the settlement history of an area. Methods based on the serial coalescent algorithm have been developed to test whether the population continuity hypothesis can be statistically rejected by analysing DNA samples from the same region but of different ages. Rejection of this hypothesis is indicative of a large genetic shift, possibly due to immigration occurring between two sampling times. However, this approach is only able to reject a model of full continuity model (a total absence of genetic input from outside), but admixture between local and immigrant populations may lead to partial continuity. We have recently developed a method to test for population continuity that explicitly considers the spatial and temporal dynamics of populations. Here, we extended this approach to estimate the proportion of genetic continuity between two populations, using ancient genetic samples. We applied our original approach to the question of the Neolithic transition in Central Europe. Our results confirmed the rejection of full continuity, but our approach represents an important step forward by estimating the relative contribution of immigrant farmers and of local hunter-gatherers to the final Central European Neolithic genetic pool. Furthermore, we show that a substantial proportion of genes brought by the farmers in this region were assimilated from other hunter-gatherer populations along the way from Anatolia, which was not detectable by previous continuity tests. Our approach is also able to jointly estimate demographic parameters, as we show here by finding both low density and low migration rate for pre-Neolithic hunter-gatherers. It provides a useful tool for the analysis of the numerous ancient DNA data sets that are currently being produced for many different species.

Investigating the sex-related geometric variation of the human cranium.

Accurate sexing methods are of great importance in forensic anthropology since sex assessment is among the principal tasks when examining human skeletal remains. The present study explores a novel approach in assessing the most accurate metric traits of the human cranium for sex estimation based on 80 ectocranial landmarks from 176 modern individuals of known age and sex from the Athens Collection. The purpose of the study is to identify those distance and angle measurements that can be most effectively used in sex assessment. Three-dimensional landmark coordinates were digitized with a Microscribe 3DX and analyzed in GNU Octave. An iterative linear discriminant analysis of all possible combinations of landmarks was performed for each unique set of the 3160 distances and 246,480 angles. Cross-validated correct classification as well as multivariate DFA on top performing variables reported 13 craniometric distances with over 85% classification accuracy, 7 angles over 78%, as well as certain multivariate combinations yielding over 95%. Linear regression of these variables with the centroid size was used to assess their relation to the size of the cranium. In contrast to the use of generalized procrustes analysis (GPA) and principal component analysis (PCA), which constitute the common analytical work flow for such data, our method, although computational intensive, produced easily applicable discriminant functions of high accuracy, while at the same time explored the maximum of cranial variability.

Age-related changes in the craniofacial region in a modern Greek population sample of known age and sex.

The aim of this paper is to investigate ageing changes in craniofacial region in both sexes and evaluate whether these shape changes are substantial to achieve age discrimination of samples used in anthropological analyses. The study sample consisted of 157 crania of known sex and age (81 males and 76 females) belonging to individuals who lived in Greece during the twentieth century. The sample was divided in three age groups: young adults (YA, 18-39 years old), middle adults (MA, 40-59 years old) and old adults (OA, >60 years old). The three-dimensional coordinates of 31 ecto-cranial landmarks were digitized using a Microscribe 3DX contact digitizer, and landmark configurations were analyzed using the generalized least-squares Procrustes method. The results indicate that both males and females show significant difference among the age groups; however, shape differences can not be used for age group discrimination due to a large range on the accuracy of age group classification. The morphometric changes related to age were different between sexes.

Cranium asymmetry in a modern Greek population sample of known age and sex.

The aim of this paper is to evaluate and quantify cranium asymmetry, sexual differences in the set of individual asymmetry scores, and the relationship between fluctuating asymmetry and age, in a modern Greek population sample. In addition, we test for the developmental origins of health and disease hypothesis by assessing the correlation between fluctuating asymmetry and cause of death. The study sample consisted of 173 crania of known sex and adult age (92 males, 81 females) belonging to individuals who lived in Greece during the twentieth century. The three-dimensional coordinates of 77 ectocranial landmarks were digitized using a Microscribe 3DX contact digitizer and landmark configurations were analyzed using the generalized least-squares Procrustes method. Regarding directional asymmetry, the results show that the human skull has a tendency for a left-side excess for the Greek population. No significant directional asymmetry differences between the sexes are found. The highest levels of fluctuating asymmetry for both sexes are located on the skull base. The levels of fluctuating asymmetry in all cranial regions appear higher for males than females. Nevertheless, these differences do not present any statistical significance between sexes. Additionally, there is no relationship between fluctuating asymmetry scores and age for both males and females. Finally, the results of this study could not confirm that early development has a significant impact on adult health outcomes.

Multidisciplinary Identification of the Controversial Freedom Fighter Jörg Jenatsch, Assassinated 1639 in Chur, Switzerland.

Jörg Jenatsch, a leading freedom fighter during the Thirty Year's War in Graubünden, Switzerland, was assassinated on carnival 1639. Jenatsch's controversial biography and the unclear circumstances of his death inspired the formation of various legends, novels and films. In 1959, a skeleton discovered in the cathedral of Chur with remains of wealthy baroque clothing was tentatively attributed to Jenatsch. Here, we reassess the skeleton based on a new exhumation. Our multidisciplinary analysis and the head injuries are consistent with reports of the eyewitnesses of the crime, demonstrating that Jenatsch was killed from behind with a semi-sharp implement, supposedly an axe, as well as by a blow with a broad-surfaced object. Moreover, our facial reconstruction closely matches an oil portrait of Jenatsch, and the HIrisPlex system applied to DNA-extracts from the femoral bone reveals brown eye and dark brown hair colour, which coincides well with the portrait, too. Finally, isotope analysis of the femoral bone and a molar support Jenatsch's high social status, luxury diet and a high mobility in the last decade of his life. This multidisciplinary approach thus reinforces personal identification and provides additional insight into the life of this important historic person beyond written resources.

Early farmers from across Europe directly descended from Neolithic Aegeans.

Farming and sedentism first appeared in southwestern Asia during the early Holocene and later spread to neighboring regions, including Europe, along multiple dispersal routes. Conspicuous uncertainties remain about the relative roles of migration, cultural diffusion, and admixture with local foragers in the early Neolithization of Europe. Here we present paleogenomic data for five Neolithic individuals from northern Greece and northwestern Turkey spanning the time and region of the earliest spread of farming into Europe. We use a novel approach to recalibrate raw reads and call genotypes from ancient DNA and observe striking genetic similarity both among Aegean early farmers and with those from across Europe. Our study demonstrates a direct genetic link between Mediterranean and Central European early farmers and those of Greece and Anatolia, extending the European Neolithic migratory chain all the way back to southwestern Asia.

Modeling ancient Egyptian mummification on fresh human tissue: macroscopic and histological aspects.

Many studies have been concerned with the ancient Egyptian mummification method; nevertheless, little effort has been made to explore it experimentally. The goal of this study is to apply evidence-based diagnostic criteria and state-of-the art methodology in order to improve knowledge on soft tissues preservation and postmortem alterations. Two human lower limbs (LL) from a female donor were (1) "naturally" mummified by dry heat and (2) artificially in natron. At specific time intervals a macroscopic and radiological examination of the LL was performed and skin and muscle samples were taken for histological and biomolecular analysis. Temperature, humidity, pH, and weight of the LL were systematically measured. The mummification by dry heat was stopped after 7 days due to unexpected lack of mummification progress. The mummification in natron was completed successfully after 208 days. The humidity, the external temperature, and the pH were proven with Pearson correlation and principal component analysis as important factors for the mummification process. The steady removal of water from the tissues through the natron has prevented the putrefaction. This is also evident in the absence of bacteria or fungi through the microbiological analysis. The histological analysis revealed very good preservation of the skin and the muscle tissues. In the muscular sample certain degree of structural disintegration can be seen, particularly affecting the epimysium whilst in the skin samples the epidermis, especially the stratum corneum, is mostly affected. The samples show better preservation compared with ancient Egyptian sections and other mummified tissues from historic or forensic context.

Histology of a Woolly Mammoth (Mammuthus primigenius) Preserved in Permafrost, Yamal Peninsula, Northwest Siberia.

In 2007, the baby woolly mammoth (Mammuthus primigenius) named Lyuba was found frozen in the Siberian tundra permafrost along the Yuribey River. She was proclaimed the best-preserved mammoth discovery. As part of the endoscopic examination of Lyuba, tissue samples of hair, muscle, and internal organs were taken. The sectioned biopsies were stained using standard and special histological stains. In general, the microscopic preservation of the tissue was good although no clearly identifiable cell nuclei were found by standard staining methods. Only a few cell nuclei could be identified in some samples when fluorescence stained with DAPI. The best-preserved structures were collagen fibers and muscle tissue, which gave some structural resemblance to the organs. In the hairs, evidence of pigmentation, a scaly surface, diagonal intra-hair structures, and a medulla were seen. Fat droplets could be identified with Sudan Red in the subcutaneous fat sample and in several organs. Bacteria were seen on the lumen side of the small intestine and caecum, and in the liver and lung tissue. In addition, fungi and pollen were seen in the lung sample. In the wall of the caecum and small intestine, blood vessels and nerves were visualized. Iron was identified in the vivianite sample. Some biopsies compared well structurally with the African elephant tissue sections. The histological findings support the theory that Lyuba drowned in muddy water. The microscopic tissue preservation and cell nuclei destruction indicate that Lyuba's body underwent at least one freeze-thaw cycle.

Post mortem DNA degradation of human tissue experimentally mummified in salt.

Mummified human tissues are of great interest in forensics and biomolecular archaeology. The aim of this study was to analyse post mortem DNA alterations in soft tissues in order to improve our knowledge of the patterns of DNA degradation that occur during salt mummification. In this study, the lower limb of a female human donor was amputated within 24 h post mortem and mummified using a process designed to simulate the salt dehydration phase of natural or artificial mummification. Skin and skeletal muscle were sampled at multiple time points over a period of 322 days and subjected to genetic analysis. Patterns of genomic fragmentation, miscoding lesions, and overall DNA degradation in both nuclear and mitochondrial DNA was assessed by different methods: gel electrophoresis, multiplex comparative autosomal STR length amplification, cloning and sequence analysis, and PCR amplification of different fragment sizes using a damage sensitive recombinant polymerase. The study outcome reveals a very good level of DNA preservation in salt mummified tissues over the course of the experiment, with an overall slower rate of DNA fragmentation in skin compared to muscle.

Y-chromosomal analysis identifies the skeletal remains of Swiss national hero Jörg Jenatsch (1596-1639).

Jörg Jenatsch was a Swiss defender of independence and a fighter for liberty in the 17th century. With the help of three living male members of the Jenatsch family, we successfully identified a skeleton exhumed from Chur cathedral as the remains of Jörg Jenatsch. Our conclusion was based upon complete Y-STR and Y-SNP profiles that could be generated by replicate analyses of a bone sample available to us. The skeleton and the three living family members carried the same Y-SNP haplogroup, but were discordant at three of 23 Y-STR loci. This notwithstanding, conservative biostatistical evaluation of the data suggests that the Chur skeleton is at least 20 times more likely than not to be Jörg Jenatsch.

"Modeling ancient Egyptian embalming": radiological assessment of experimentally mummified human tissue by CT and MRI.

OBJECTIVE: To assess changes in different tissues during the process of artificial mummification by natron using computed tomography (CT) and magnetic resonance imaging (MRI), and to translate the results to image interpretation in paleoradiological studies of ancient mummies. MATERIALS AND METHODS: A human lower limb (LL) was amputated from a female donor 24 h post-mortem and mummified by artificial natron (54 % NaCl, 16 % Na2SO4, 18 % Na2CO3 12 % NaHCO3) in ancient Egyptian style. The LL was kept in a fume hood at 16-25 °C and 30-75 % relative humidity. CT and MRI were performed at specific intervals with quantitative evaluation of Hounsfield units (HU) and signal intensities (SI). RESULTS: Evaluated tissues showed different HU and SI changes during the experimental mummification. All tissues revealed an overall but varying increase of HU in CT examinations. All tissues except for the compact bone revealed an overall but varying decrease of SI in the IR and T2-weighted sequences of the MRI. Typical findings included a distinct increase of HU in the cutis at the end of the study and a temporary increase of SI in the IR and T2-weighted sequences in all muscle groups. CONCLUSIONS: Radiological findings showed a regular, controlled and effective dehydration by the applied natron without detectable putrefaction. Evaluated tissues revealed different radiological changes during the experiment, which altogether led to preservation of the tissues without radiologically identifiable destruction. The cutis revealed radiological signs of direct interaction with the natron in the form of covering and possibly permeation.