Carbon, nitrogen, and sulfur elemental and isotopic variations in mouse hair and bone collagen during short-term graded calorie restriction.

This study characterized the effect of calorie restriction (CR) on elemental content and stable isotope ratio measurements of bone "collagen" and hair keratin. Adult mice on graded CR (10-40%; 84 days) showed decreased hair δ 15N, δ 13C, and δ 34S values (significantly for δ 15N) with increasing CR, alongside a significant increase in bone "collagen" δ 15N values and a decrease in "collagen" δ 13C values. We propose this was likely due to the intensified mobilization of endogenous proteins, as well as lipids in newly synthesized "collagen". Elemental analysis of bone "collagen" revealed decreased carbon, nitrogen, and sulfur % content with increasing CR which is attributed to a change in the in vivo bone "collagen" structure with extent of CR. This complexity challenges the use of elemental indicators in the assessment of collagen quality in archaeological studies where nutritional stress may be a factor.

No Bones About It: Sex Is Binary.

Anthropologists have led the way in formulating techniques that reveal skeletal differences between males and females. Understanding of physical differences in the pelvis related to childbirth, hormonal impacts on bones, and extensive comparative studies have provided anthropologists with an array of traits and measurements that help them estimate sex using just bones. Forensic anthropologists and bioarcheologists are improving their ability to differentiate males and females by increasing research on a variety of postcranial bones and through the use of molecular data, especially new methods called proteomics, to identify sex in prepubescent juveniles. As remains from more cultures and time periods are studied, sex identification will continue to improve, because skeletal sex differences are in large part biologically determined. Yet, anthropologists have also been at the forefront of arguing that sex lies on a spectrum. Anthropologists who view sex as on a spectrum may deter sex identification progress; from their perspective, an individual of an undetermined sex may just be a nonbinary individual. Anthropologists who consider sex is on a spectrum are coming to this conclusion in part because they are looking for anatomical ideals, mistaking pathology for variation, and confusing independent variables with dependent variables. Nonetheless, anthropologists need to continue to improve sex identification techniques to reconstruct the past accurately, which may reveal less strict sex roles than previously presumed and help with the identification of crime victims. Forensic anthropologists should also increase their efforts to identify whether individuals have undergone medical procedures intended to change one's gender due to the current rise in transitioning individuals.

Fatty Acids and Starch Identification within Minute Archaeological Fragments: Qualitative Investigation for Assessing Feasibility.

Within the realm of archaeology, the analysis of biomolecules assumes significant importance in elucidating historical dietary patterns and their implications for contemporary contexts. To achieve this, knowledge and tools of both chemistry and archaeology are essential to yield objective outcomes and conduct analyses of archaeological materials for the detection of biomolecules. Usually, only minuscule remnants of ceramic fragments are retrieved from excavations, which limits the feasibility of comprehensive laboratory analysis. This study aimed to establish a protocol for analyzing fatty acids and starch from archaeological food utensils with minimal sample quantities. Various experiments were conducted to replicate preparations that might have occurred in archaeological vessels, aiming to establish the optimal protocol. The analyses were performed using clay griddles, subjecting vegetable oil to varying temperatures for fatty acid assessment. For starch analysis, a series of experiments encompassed diverse forms of potato preparations (pulp, chuño, tortilla, carbonization, and freeze-drying) and maize (flour, tortilla, and carbonization). The verification of the experiments was confirmed by conducting identical analyses, as developed in the current study, on authentic archaeological fragments. The principal outcomes of this investigation include the successful extraction of both types of biomolecules using only 0.25 g of the sample, obtained through direct scraping from the vessel. Soxhlet extraction was identified as the most efficient strategy to recover fatty acids. Additionally, a comprehensive protocol for the identification of starch extraction was developed. This study has, for the first time, elucidated two detailed methodologies for the extraction of fatty acids and starch in scenarios in which researchers can obtain limited quantities of archaeological food utensil fragments.

Fecal biomarkers and micromorphological indicators of sheep penning and flooring at La Fortaleza pre-Hispanic site (Gran Canaria, Canary Islands).

This study explores the lipid content and micromorphological features of sediment samples from two dwelling structures at the pre-Hispanic site of La Fortaleza in Santa Lucía de Tirajana (Gran Canaria, Spain). Previous field identification of possible sedimentary excrements inside the dwellings motivated chromatographic fecal biomarker analysis and micromorphology. The micromorphological samples reveal a complex dung-rich stratified sequence involving different layers of mixed composition, including reworked dung, clay, wood ash, and domestic refuse. The results of the lipid analysis corroborate the fecal nature of the sample and indicate the source animal: sheep. Coupled with the field evidence, the data suggest that the deposit is anthropogenic and represents a sequence of floor foundations, dung floors, and domestic and architectural refuse. This study provides valuable taxonomic and site use data for the understanding of the aboriginal societies of the Canary Islands and shows the efficacy of combining field observations with high-resolution geoarchaeological methods.

Microstratigraphic, lipid biomarker and stable isotope study of a middle Palaeolithic combustion feature from Axlor, Spain.

Archaeological research has increasingly focused on studying combustion features as valuable sources of information regarding past technological and cultural aspects. The use of microstratigraphic and biomolecular techniques enables the identification of combustion residues and substrate components, and infer about past fire-related activities and the environments. Our study conducted on a combustion feature (Level N, ∼100 Ka) at the Axlor cave, a Middle Paleolithic site in northern Iberia, exemplifies the interdisciplinary approach to combustion features. Micromorphological features revealed depositional activities associated with occupations such as hearth rake-out and trampling. Through molecular (n-alkanes, n-alcohols, and n-fatty acids) and isotopic analysis (δ13C16:0 and δ13C18:0), we infer the good preservation of organic matter, the contributions of non-ruminant fats, and the dead-wood gathering strategies by Neanderthal groups. By combining microstratigraphic and biomolecular approaches, our study significantly contributes to the advancement of our current understanding of Neanderthal pyrotechnology.

Efficiency of Portable X-ray Fluorescence for Distinguishing Lead Slag.

In the Minier Valley (France, Aveyron), an archaeological site excavation has led to the discovery of 25 kg of lead (Pb) slag, found in a secondary position. On a macroscopic level, these slags can be classified into two distinct categories, vitreous black and porous gray. This clear visual differentiation has been previously documented at other Pb-silver (Ag) metallurgy sites dating from the 12th century to the 14th century in Europe, and more particularly in the workshops of Mont Lozère, located approximately 80 km away from Le Minier. However, to date, scant evidence exists regarding the specific metallurgical process that produced this slag. This study presents the results of the analysis of 218 Pb slags (Supplemental Material) using portable X-ray fluorescence (pXRF). This method does not appear to be ideally suited for investigating this type of artifact, as it does not provide quantification of oxygen content. Nevertheless, it can be used to quickly identify trends and determine whether the two identified facies exhibit dissimilar chemical compositions. The results indicate that, based on the elemental compositions of the slags, no discernible distinction can be made to categorize them into two separate groups, both in bivariate and multivariate analyses. Consequently, it can be inferred that the gray and black slags originate from a single process executed within the same type of furnace. Scanning electron microscopy energy dispersive spectrometry analyses on selected samples demonstrate that variations in metal content are considerably exaggerated by pXRF, but that the trends observed are confirmed. The use of pXRF is therefore a relevant tool for determining whether or not Pb-Ag slag is the result of the same process. However, this initial approach needs to be refined by other analyses to confirm whether the smelted ores are of local origin and to be able to better define the details of the metallurgical process used.

Age-depth model for uppermost Ndutu Beds constrains Middle Stone Age technology and climate-induced paleoenvironmental changes at Olduvai Gorge (Tanzania).

Olduvai Gorge in northern Tanzania is part of a globally important archeological and paleoanthropological World Heritage Site location critical to our understanding of modern human evolution. The Ndutu Beds in the upper part of the geological sequence at Olduvai Gorge represent the oldest unit to yield modern Homo sapiens skeletal material and Middle Stone Age technology. However, the timing of the deposition of the Ndutu Beds is poorly constrained at present, which limits our understanding of the paleoenvironments critical for contextualizing H. sapiens and related technologies in the Olduvai Basin. Using a suite of 15 luminescence ages of sedimentary core samples, combined with Bayesian statistics, this study provides a new higher-resolution age-depth model for the deposition of the uppermost Upper Ndutu and Naisiuiu Beds cored by the Olduvai Gorge Coring Project. The luminescence and modeled ages are presented as ±1 σ uncertainties. The Ndutu Beds intersected by the Olduvai Gorge Coring Project cores are dated to between 117.1 ± 17.9 and 45.3 ± 4.2 ka (between 125.9 ± 26.5 and 45.8 ± 8.2 ka modeled ages), while a probable overlying layer of Naisiusiu Beds dates to 23.7 ± 10.9 to 12.1 ± 1.7 ka (25.7 ± 18.9 ka and 12.0 ± 3.4 ka modeled age). Time-averaged accretion rates are derived during this time: (1) initially low rates (<5 cm ka-1) from the bottom of the core at 117.1 ± 17.9 ka up to 95.3 ± 11.1 ka (125.9 ± 26.5 to 95.5 ± 23.3 ka modeled ages); (2) the middle section spanning between 95.3 ± 11.1 and 62.7 ± 5.7 ka (95.5 ± 23.3 to 61.9 ± 10.4 ka modeled ages) with mean rates above 15 cm ka-1; and (3) the last 62.7 ± 5.7 ka (61.9 ± 10.4 ka modeled age) where the accretion rate reduces to below 5 cm ka-1. This reduction can be explained by the evolution of the gorge system that was likely driven by subsidence of the Olbalbal depression and changes in climate, particularly precipitation and resulting lake and base level changes. Older Upper Ndutu and Lower Ndutu Beds are contained within proto-gorges within the modern gorge system.

Use of strontium isotope ratios in potential geolocation of Ajnala skeletal remains: a forensic archeological study.

Stable isotope methods for provenance of unidentified human remains are relatively a newer field of enquiry in forensic archeology. It is of great interest for forensic experts these days. The application of strontium isotope analyses for estimating geolocation of archeological remains is of great interest in bioarcheology and modern forensics. The strontium (Sr) isotope composition of human bones and teeth has been widely used to reconstruct an individual's geo-affiliation, residential mobility, and migration history. Thousands of unknown human remains, reportedly belonging to 282 Indian soldiers of 26th Native Bengal regiment and killed in 1857, were exhumed non-scientifically from an abandoned well situated underneath a religious structure at Ajnala (Amritsar, India). Whether these remains belonged to the individuals, local or non-local to the site, was the important forensic archeological question to be answered by doing their thorough forensic anthropological examinations. In the present study, 27 mandibular teeth (18 s molars, 6 first molars, and 3 premolars) collected from the Ajnala skeletal assemblage were processed for strontium isotope analysis, and the measured ratios were compared with published isotope baseline data to estimate the locality status of these remains. The Sr isotopic values were concentrated in the range of 0.7175 to 0.7270. The comparative analysis of isotopic ratios revealed that most individuals buried in the Ajnala well have 87Sr/86Sr values close to the river as well as groundwater of the Gangetic plain (less radiogenic 87Sr/86Sr ~ 0.716); most likely originated near Varanasi (Uttar Pradesh, India) region, whereas the individuals with higher 87Sr/86Sr ratios (~ 0.7200) probably resided in the West Bengal and Bihar areas where the river as well as groundwater of the Gangetic plain is relatively more radiogenic. Thus, the strontium isotope results reveal that the Ajnala individuals did not grow up or live in the Amritsar region during their childhood, and this observation complemented the previous forensic anthropological and molecular findings. There is very little Indian data on the bioavailable strontium, so the inferences from the present study estimating Sr isotope abundances are expected to provide baseline data for future forensic provenance studies that will contribute to the global efforts of mapping Sr isotope variations by the isotope community.

An individual with Sarmatian-related ancestry in Roman Britain.

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

Towards correlative archaeology of the human mind.

Retracing human cognitive origins started out at the systems level with the top-down interpretation of archaeological records spanning from man-made artifacts to endocasts of ancient skulls. With emerging evolutionary genetics and organoid technologies, it is now possible to deconstruct evolutionary processes on a molecular/cellular level from the bottom-up by functionally testing archaic alleles in experimental models. The current challenge is to complement these approaches with novel strategies that allow a holistic reconstruction of evolutionary patterns across human cognitive domains. We argue that computational neuroarcheology can provide such a critical mesoscale framework at the brain network-level, linking molecular/cellular (bottom-up) to systems (top-down) level data for the correlative archeology of the human mind.

Review: Computational analysis of human skeletal remains in ancient DNA and forensic genetics.

Degraded DNA is used to answer questions in the fields of ancient DNA (aDNA) and forensic genetics. While aDNA studies typically center around human evolution and past history, and forensic genetics is often more concerned with identifying a specific individual, scientists in both fields face similar challenges. The overlap in source material has prompted periodic discussions and studies on the advantages of collaboration between fields toward mutually beneficial methodological advancements. However, most have been centered around wet laboratory methods (sampling, DNA extraction, library preparation, etc.). In this review, we focus on the computational side of the analytical workflow. We discuss limitations and considerations to consider when working with degraded DNA. We hope this review provides a framework to researchers new to computational workflows for how to think about analyzing highly degraded DNA and prompts an increase of collaboration between the forensic genetics and aDNA fields.

Ancestry and kinship in a Late Antiquity-Early Middle Ages cemetery in the Eastern Italian Alps.

In South Tyrol (Eastern Italian Alps), during Late Antiquity-Early Middle Ages, archeological records indicate cultural hybridization among alpine groups and peoples of various origin. Using paleogenomics, we reconstructed the ancestry of 20 individuals (4th-7th cent. AD) from a cemetery to analyze whether they had heterogeneous or homogeneous ancestry and to study their social organization. The results revealed a primary genetic ancestry from southern Europe and additional ancestries from south-western, western, and northern Europe, suggesting that cultural hybridization was accompanied by complex genetic admixture. Kinship analyses found no genetic relatedness between the only two individuals buried with grave goods. Instead, a father-son pair was discovered in one multiple grave, together with unrelated individuals and one possible non-local female. These genetic findings indicate the presence of a high social status familia, which is supported by the cultural materials and the proximity of the grave to the most sacred area of the church.

A multidisciplinary overview on the Tupi-speaking people expansion.

The cultural and biological diversity of South American indigenous groups represent extremes of human variability, exhibiting one of the highest linguistic diversities alongside a remarkably low within-population genetic variation and an extremely high inter-population genetic differentiation. On top of that, this region has seen some of the most dramatic demographic events in human history unleashed by the European colonization of the Americas. As a result of this process, the distribution of indigenous populations has been radically changed. In this review we focus on the Tupi, the largest and most widespread linguistic family in eastern South America. Tupi are believed to have originated in southwestern Amazon, from where some of its subfamilies expanded into other parts of the Amazon and, in the case of the Tupi-Guarani, beyond its borders. Recent evidence from archaeology, linguistics, and genetics aligns with José Brochado's Tupi Expansion model. He proposed that the gradual development of agricultural systems within the Amazon resulted in population growth and, eventually, territorial expansion. This model also supports separate Tupi Expansion branches: Tupinambá (Atlantic coast) and Guarani (south, midwest Brazil). Although being the most populous group on Brazil's Atlantic coast, which was the most affected by European colonization, the Tupi still account for roughly 20% of the country's overall indigenous population. Finally, despite its importance and more than a century of research on the Tupi and their expansion history, many key questions remain unanswered, which we attempt to summarize and explore here.

A diversidade cultural e biológica dos grupos indígenas sul-americanos representam extremos da variabilidade humana, abrangendo uma das maiores diversidades linguísticas e uma das menores diversidades genéticas intrapopulacionais do mundo, assim como uma das maiores diferenciações genéticas interpopulacionais. Além disso, esta região presenciou alguns dos eventos demográficos mais dramáticos da história da humanidade, desencadeados pela colonização européia das Américas. Como resultado desse processo, a distribuição das populações indígenas mudou radicalmente. Nesta revisão, focamos nos Tupi, o maior e mais difundido grupo linguístico do leste da América do Sul, dentre as sete principais famílias linguísticas do continente. Acredita-se que os Tupi tenham se originado no sudeste da Amazônia, de onde algumas de suas subfamílias se expandiram para outras partes da Amazônia e, no caso dos Tupi-Guarani, para além de suas fronteiras. Evidências recentes da arqueologia, linguística e genética alinham-se com o modelo de Expansão Tupi de José Brochado. Ele propôs que o desenvolvimento gradual dos sistemas agrícolas na Amazônia resultou no crescimento populacional e, eventualmente, na expansão territorial. Esse modelo também apóia ramos separados da Expansão Tupi: Tupinambá (costa atlântica) e Guarani (sul e centro-oeste do Brasil). Embora sejam o grupo mais populoso da costa atlântica do Brasil, que foi a mais afetada pela colonização europeia, os Tupi ainda representam cerca de 20% da população indígena total do país. Finalmente, apesar de sua importância e de mais de um século de pesquisa sobre os Tupi e sua história de expansão, muitas questões-chave permanecem sem solução, as quais tentamos resumir e explorar aqui.

Curated cauldrons: Preserved proteins from early copper-alloy vessels illuminate feasting practices in the Caucasian steppe.

Large metal and metal-alloy cauldrons first appear on the far western steppe and Caucasus region during the Maykop period (3700-2900 BCE); however, the types of foods or beverages cooked in and served from these vessels have remained mysterious. Here, we present proteomic analysis of nine residues from copper-alloy cauldrons from Maykop burial contexts where we identify muscle, blood, and milk proteins specific to domesticated, and possibly wild, ruminants. This study clearly demonstrates that the earliest, large-volume feasting vessels contained both primary and secondary animal products, likely prepared in the form of a stew.

The early lives of the islanders: Stable isotope analysis of incremental dentine collagen from the prehispanic period of the Canary Islands.

OBJECTIVES: This study presents isotopic information for incremental dentine collagen and bone bulk collagen from individuals from the Canary Islands (Tenerife and Gran Canaria) to explore dietary differences during childhood life. MATERIALS AND METHODS: Eight individuals have been studied, which comprises 122 δ15 N and δ13 C incremental dentine measurements and eight bulk bone collagen analyses. A baseline of potentially consumed food sources has been developed for comparative purposes. A food reconstruction using isotopic transferred signals (FRUITS) model of probable contributions of each food source towards the diet of each individual has been developed. All samples but one belongs to the later period of indigenous occupation of the archipelago. RESULTS: The dentine collagen data are presented in correlated δ13 C and δ15 N plots per individual, showing the isotopic changes throughout time. δ15 N values for each individual tend to be variable whereas δ13 C data are generally more stable with a range of +9.1 to +14‰ for δ15 N and -17.4 to -20.8‰ for δ13 C. CONCLUSION: The isotopic analysis allows for the reconstruction of eight dietary profiles, which allow us to estimate the different dietary protein sources. The FRUITS model shows different percentages of the primary food sources for each individual. Where both δ13 C and δ15 N are elevated, this could be indicative of a higher marine contribution to the diet. There appear to be two main dietary profiles identifiable in the dataset and these may be related to changes in status or place of residence. Short-term variations in δ13 C and δ15 N and opposing co-variance of isotopic values can be indicative of nutritional stress, although metabolic changes during growth are also considered.

Bioarchaeological analyses reveal long-lasting continuity at the periphery of the Late Antique Roman Empire.

The Basel-Waisenhaus burial community (Switzerland) has been traditionally interpreted as immigrated Alamans because of the location and dating of the burial ground - despite the typical late Roman funeral practices. To evaluate this hypothesis, multi-isotope and aDNA analyses were conducted on the eleven individuals buried there. The results show that the burial ground was occupied around AD 400 by people belonging largely to one family, whereas isotope and genetic records most probably point toward a regionally organized and indigenous, instead of an immigrated, community. This strengthens the recently advanced assumption that the withdrawal of the Upper Germanic-Rhaetian limes after the "Crisis of the Third Century AD" was not necessarily related to a replacement of the local population by immigrated Alamannic peoples, suggesting a long-lasting continuity of occupation at the Roman periphery at the Upper and High Rhine region.

Improving the extraction of ancient Yersinia pestis genomes from the dental pulp.

Ancient DNA preserved in the dental pulp offers the opportunity to characterize the genome of some of the deadliest pathogens in human history. However, while DNA capture technologies help, focus sequencing efforts, and therefore, reduce experimental costs, the recovery of ancient pathogen DNA remains challenging. Here, we tracked the kinetics of ancient Yersinia pestis DNA release in solution during a pre-digestion of the dental pulp. We found that most of the ancient Y. pestis DNA is released within 60 min at 37°C in our experimental conditions. We recommend a simple pre-digestion as an economical procedure to obtain extracts enriched in ancient pathogen DNA, as longer digestion times release other types of templates, including host DNA. Combining this procedure with DNA capture, we characterized the genome sequences of 12 ancient Y. pestis bacteria from France dating to the second pandemic outbreaks of the 17th and 18th centuries Common Era.

Volatile organic compounds (VOCs) as a rapid means for assessing the source of coprolites.

The odor of rehydrated coprolites can be used as an informal means of fecal identification. To date, the analysis of volatiles emitted by coprolites from different sources has not been attempted, and the possibility of utilizing volatile organic compounds (VOCs) as fecal biomarkers unexplored. VOCs released by coprolites from the Paisley Caves, were analyzed using solid-phase microextraction (SPME), to assess the variance of results from different coprolites (carnivores, herbivores, or humans). Coprolites from carnivores can be clearly distinguished from those produced by herbivores and humans; these latter two are separated to a lesser degree. Eight discriminatory compounds differentiated between the coprolite sources, and their identities were verified using reference standards. Coprolites and their associated sediments could not be differentiated between using this method, suggesting leaching of VOCs into the burial matrix. This work provides an alternative, more rapid way to assess coprolite origin.

The detection of clandestine graves in an arid environment using thermal imaging deployed from an unmanned aerial vehicle.

The Middle East is one of the world regions that has frequently suffered from armed conflicts that resulted in mass burials. However, the detection of clandestine graves in such an arid environment by deploying remote sensing payload on unmanned aerial vehicles (UAVs) has received little attention. The present study used a UAV equipped with a thermal sensor aimed at narrowing down the search area of possible gravesites in the arid climate of Kuwait. The enclosed research area, which includes both control and experimental mass graves, was imaged for 18 months. The variation in topsoil temperature and soil moisture between the graves and their surroundings was evaluated. The results of the analysis demonstrated the effectiveness of thermal imaging techniques in detecting heat produced from buried sheep carcasses and detecting the change in grave soil moisture for our research environment for 7 and 10 months, respectively. The buried animals significantly influenced the topsoil temperature (p = 0.044), while the height from which the images were captured had an insignificant effect on the measured temperature within the range tested (p = 0.985). Furthermore, there was a negative correlation (-0.359) between grave temperature and the calculated soil moisture. The results from these cost- and time-effective search methods presented in this study confirm their potential for the detection of burial sites in an arid environment.