Archaeological Skeletal Remains Research Articles

Chronic Maxillary Sinusitis: A comparison of osteological and CT methods of diagnosis

ObjectiveTo assess the differences between endoscopic and radiological methods of analysis for diagnosing chronic maxillary sinusitis (CMS) in archaeological skeletal remains. Materials32 crania from a Dutch post-medieval rural population. MethodsWe assessed the presence of bone changes indicative of CMS (i.e., bone growth and bone resorption) both endoscopically and through computed tomography (CT), and then compared results. ResultsWe observed moderate agreement between bone growth scores obtained through endoscopy and CT, and fair agreement when assessing bone resorption. ConclusionsCMS prevalence rates observed through CT may be comparable to rates assessed endoscopically, although caution is needed when making direct comparisons. SignificanceThis is the first study comparing data obtained through endoscopy and radiological methods in the study of CMS, informing paleopathologists about potential biases in data comparison. LimitationsOur small sample size likely impacted results. Suggestions for further researchFurther research is advised to fully explore the comparability of endoscopic and radiological method of analysis in the study of sinusitis. Data AvailabilityThe datasets generated and analyzed during the current study are available upon request.

Odontological analysis of medieval archaeological samples from Krešića greblje near Čitluk, Bosnia and Herzegovina - An auxiliary method to genetic analysis

Previous research on archaeological skeletal remains in Bosnia and Herzegovina rarely included odontological analysis. The objective of this research was biological profiling through odontological analysis (dental morphological characteristics, estimation of sex and dental age at death) in medieval archaeological skeletal remains samples from Krešića greblje in Paoča (Čitluk municipality, Bosnia and Herzegovina). After the excavations of the archaeological site, odontological analysis was performed before genetic analysis. The sample comprised 22 permanent teeth from 6 individuals. Odontological analysis included odontometric estimation of sex, estimation of age at death by two morphological methods, and an analysis of dental traits and dental pathology. Genetic analysis was used for sex determination. The results of odontological analysis classified all individuals as adults, with estimated age at death 20-25 in one individual, 30-35 in three individuals, 35-45 in one individual, and one individual older than 45 years of age. Odontometric sex estimation was in high concordance with sex determination by genetic analysis, and correctly classified five individuals’ sex (four females and one male). In one individual odontometric sex was assessed as female while genetic analysis revealed male. Morphological dental traits and dental pathology were investigated as well, adding to a better understanding of the odontological characteristics of the medieval population in the area of Bosnia and Herzegovina. Sex and dental age estimation given by odontological analysis provide fast and cost-effective information, and should be performed as an auxiliary before genetic analysis.

Automated DNA extraction from recent and archaeological human skeletal remains using QIAGEN's EZ2 Connect Fx Instrument – an experience report

The aim of this study was to investigate the efficiency of the automated DNA extraction method using the EZ2 Connect Fx instrument (QIAGEN), showcasing its ability to achieve swift and reliable results from bone samples. Genomic DNA extraction from 16 archaelogical and recent bone samples was performed with a commercial EZ1&2® DNA Investigator® Kit (QIAGEN). Prior to PCR amplification, DNA concentration was quantified using Qubit™ Fluorometer (Thermo Fisher Scientific). Amplification was carried out with the Investigator 24plex Kit (QIAGEN). DNA profiles were generated using 3500 Genetic Analyzer (Applied Biosystems) and analysed with GeneMapper™ ID-X 1.6 Software (Applied Biosystems). For nine archaeological dental samples, eight exhibited partial profiles, and one showed no amplified loci. Among the seven recent femoral bone samples, one exhibited a partial profile, while six presented complete DNA profiles. In this study, we investigated the benefits of automated DNA extraction, particularly its ability to generate timely and reliable results. Additionally, this method reduces pipette and tip usage, almost completely minimising the risk of human error, exogenous DNA contamination and cross-contamination. Such efficiency is of paramount importance, especially when working with skeletal remains, namely archaeological skeletal remains that pose a challenge for molecular genetic analyses.

Mineralization and thickness of the condylar cortex in skeletal remains of children’s mandibles: A preliminary study

ObjectiveTo explore the relationship between the volumetric bone mineral density (vBMD), the thickness of the condylar cortex (Tcortex) and the hemimandibular volumes (Vhemimandible) of symmetrical and asymmetrical mandibles of children. DesignThe data collection consisted of 92 archeological skeletal remains of children’s mandibles between 1 and 12 years old. The mandibles were digitalized with a computed tomography (CT) scan, and three dimensional models were obtained. Vhemimandible was calculated using the optimal symmetry plane. The volumes were used to calculate the asymmetry index (AI). Mandibles with an AI of ≥ 3% (N = 9) and a sample of the most symmetrical mandibles (N = 9) were selected for this research. Three groups were created: a symmetrical, an asymmetrical and a pooled group. Micro-CT was used to measure the vBMD and Tcortex in four volumes of interest. The AI was calculated for these parameters as well. ResultsSignificant correlations were found between the vBMD and the Tcortex in the pooled group (P < .01) and between the AI of the vBMD and the AI of the Tcortex in the pooled (P < .01) and symmetrical group (P < .05). No significant correlations were found between the vBMD and the Vhemimandible and between the respective AIs. Between the Tcortex and the Vhemimandible a significant correlation was found in the pooled and asymmetrical group. ConclusionThere is a relationship between the vBMD and the Tcortex. The correlations between the Tcortex and the Vhemimandible are insufficient to draw firm conclusions. A relationship between the vBMD and Vhemimandible was not confirmed in this study.

Comparing extraction method efficiency for high-throughput palaeoproteomic bone species identification

High-throughput proteomic analysis of archaeological skeletal remains provides information about past fauna community compositions and species dispersals in time and space. Archaeological skeletal remains are a finite resource, however, and therefore it becomes relevant to optimize methods of skeletal proteome extraction. Ancient proteins in bone specimens can be highly degraded and consequently, extraction methods for well-preserved or modern bone might be unsuitable for the processing of highly degraded skeletal proteomes. In this study, we compared six proteomic extraction methods on Late Pleistocene remains with variable levels of proteome preservation. We tested the accuracy of species identification, protein sequence coverage, deamidation, and the number of post-translational modifications per method. We find striking differences in obtained proteome complexity and sequence coverage, highlighting that simple acid-insoluble proteome extraction methods perform better in highly degraded contexts. For well-preserved specimens, the approach using EDTA demineralization and protease-mix proteolysis yielded a higher number of identified peptides. The protocols presented here allowed protein extraction from ancient bone with a minimum number of working steps and equipment and yielded protein extracts within three working days. We expect further development along this route to benefit large-scale screening applications of relevance to archaeological and human evolution research.

Differentiating present-day from ancient bones by vibrational spectroscopy upon acetic acid treatment

Acetic acid treatment for an accurate differentiation between ancient and recent human bones was assessed using Raman and FTIR-ATR spectroscopies. Each set of skeletal samples was analysed by these techniques, prior and after chemical washing, in order to determine the variations in bone´s chemical composition and crystallinity. Bone samples were collected from several independent sources: recent bones burned under controlled experimental conditions or cremated, and archaeological (XVII century and Iron Age). The effect of acetic acid, expected to impact mostly on carbonates, was clearly evidenced in the spectra of all samples, particularly in FTIR-ATR, mainly through the bands typical of A- and B-carbonates. Furthermore, as seen for crematoria and archaeological samples, acetic acid was found to remove contaminants such as calcium hydroxide. Overall, acetic acid treatment can be an effective method for removing carbonates (exogenous but possibly also endogenous) and external contaminants from bone. However, these effects are dependent on the skeletal conditions (e.g. post-mortem interval and burning settings). In addition, this chemical washing was shown to be insufficient for an unequivocal discrimination between recent and archaeological skeletal remains. Based on the measured IR indexes, only cremated bones could be clearly distinguished.

Skeletal evidence for two external auditory canal disorder cases from medieval China

AbstractEar diseases, especially external auditory canal (EAC) disorders, are rarely reported in paleopathology. This study reports on the archaeological skeletal remains of two adults from the medieval period in China that presented with bony destruction in the external auditory canal (EAC). The observed pathological changes were evaluated macroscopic and microscopic, and micro‐CT (micro‐computed tomography) was used to examine the extent of lesion involvement. The lesions in Case 1 involved the EAC and mastoid process with smooth and regular margins. We, therefore, concluded that EAC cholesteatoma is the most likely diagnosis. In Case 2, the lesion was in the superior and posterior walls of the EAC as well as in the mastoid air cells. In this case, we arrive at the diagnosis of EAC infection. Furthermore, the potential impact of these ear diseases has been discussed. This report addresses the lack of detailed descriptions of EAC diseases in previous paleopathological publications and emphasizes the fact that the diagnosis of ear diseases in this context requires comprehensive observations of lesions as well as of all adjacent structures.

Sex determination of medieval skeletal remains

During 2019, 28 grave constructions were excavated at cemetery Metaljica (Hadzici municipality, Bosnia and Herzegovina). Archaeological excavation have revealed 17 well preserved skeletons that were subjected to anthropological, odontological and genetic analyses. To determine sex by genetic analysis, amelogenin and DYS391 loci were taken into consideration. Concordance between results of anthropological, odontological and genetic analyses applied in this research occurred in one sample out of 17. In four samples, sex was determined only by genetic analysis, since sexual dimorphism indicators were not preserved. Concordance between odontological and genetic determinations was observed in two samples, while affirmative results of anthropological and genetic analysis were obtained in only one sample. Discrepancy in the majority of samples can be atributted to the state of preservation of skeletal remains, interpopulation differences and allele drop-out. Moreover, anthropological and odontological analyses were not applicable to juvenile/subadult skeletons, since sexually dimorphic characteristics relevant for metric and morphological analysis are not developed well at that developmental period. This study emphasizes the importance of combining anthropological, odontological and genetic methods in order to determine sex of archaeological skeletal remains accurately.

Malignancy in three medieval Polish osteological collections

This paper presents the archaeological skeletal remains of four adult females presenting with cranial lesions typical of malignant neoplastic disease. The four cases were sourced from three medieval Polish skeletal assemblages: Milicz (12–13th century), Pawłów Trzebnicki (15–16th century), and Gródek nad Bugiem (13–15th century). The observed pathological skeletal changes were evaluated with a comprehensive macroscopic analysis of the skeleton, and radiography of the affected bones. The observed osteolytic lesions were largely limited to the crania and were multiple and varied in size; further internal lesions were revealed with radiography. Three cases were differentially diagnosed as highly consistent with metastatic carcinoma, and the fourth case was differentially diagnosed as typical of multiple myeloma. This report adds to the scant number of palaeopathological examples of malignant neoplasms in Poland and it discusses the possible impact of cancer on medieval lives, as well as possible factors in the occurrence of malignant neoplasms in medieval Poland.

The impact of moderate heating on human bones: an infrared and neutron spectroscopy study.

This study aims to analyse human bones exposed to low/medium temperatures (200–650°C) under experimentally controlled conditions, both oxidizing and reducing, using complementary optical and neutron vibrational spectroscopy techniques. Clear differences were observed between the aerobically and anaerobically heated bones. The organic constituents disappeared at lower temperatures for the former (ca 300°C), while they lingered for higher temperatures in anaerobic environments (ca 450–550°C). Unsaturated non-graphitizing carbon species (chars) were detected mainly for anaerobically heated samples, and cyanamide formation occurred only at 650°C in reducing settings. Overall, the main changes were observed from 300 to 400°C in anaerobic conditions and from 450 to 500°C in aerobic environments. The present results enabled the identification of specific spectroscopic biomarkers of the effect of moderate temperatures (less than or equal to 650°C) on human bone, thus contributing to a better characterization of forensic and archaeological skeletal remains subject to heating under distinct environmental settings. In particular, these data may provide information regarding cannibalism or ancient bone boiling and defleshing rituals.

Testing the cross-applicability of juvenile sex estimation from diaphyseal dimensions

Sex estimation is a crucial component of the biological profile. Stull et al. (2017) have proposed a promising juvenile sex estimation method using long bone measurements taken from a South African sample, providing relatively high classification accuracies and made easy to use via the KidStats web-based app. In this study, we test the models developed by Stull et al. (2017) on an external historic population from Lisbon, Portugal, in order to determine whether the models can be reliably applied to archeological and forensic populations outside of the original population sample. The study sample consisted of 102 individuals (45 females and 57 males) aged under 13 years at death from the Lisbon identified skeletal collection. Measurements from these individuals were used to test the flexible discriminant analysis (FDA) models given by Stull et al. (2017). Allocation accuracies were calculated for boys and girls and children over and under 2 years separately and combined. Our findings show that the models developed by Stull et al. (2017) yield poor accuracy when applied to our external population and thus can potentially be misapplied on archeological skeletal remains or forensic remains of unknown origin. A number of statistical issues may explain why models fail to be transportable or even generalizable, namely multicollinearity, model overfitting and overly optimist bootstrapped cross-validation rates. It is also likely that population differences in size and sexual size dimorphism also affected the applicability of the models. We emphasize the importance of externally validating prediction models, particularly if they are intended to be applied across populations. Our study addresses Stull and co-worker’s request for further validation of the method on populations outside of South Africa, as the models cannot be confidently applied in the field until it has been externally validated.

Allen's fossa—An attempt to dissolve the confusion of different nonmetric variants on the anterior femoral neck

AbstractA comparison of different studies is only possible if the same guidelines are used. However, due to the rather imprecise description of the different nonmetric traits on the anterior femoral neck (i.e., Allen's fossa, femoral plaque, Poirier's facet), it is impossible to compare the results of different authors. This is especially the case for the variant called Allen's fossa or cervical fossa of Allen, which has severely been mistaken in a lot of previous studies. As a consequence, a clarified and more detailed definition of this variant is required what enables the comparison of the results of different investigators. This study gives a detailed overview on the variety of different terms, which are often used simultaneously. The aim of the study is to develop a clarified definition of the term Allen's fossa, including the description of any sex‐ or age‐related differences, based on morphological examinations of archaeological skeletal remains from Karacamirli (Azerbaijan), Unterigling (Germany), and Ur (Iraq). Allen's fossa is shown to be usually bilaterally present in individuals of all age classes, including infants and rather old individuals, without any signs of modifications with increasing age. In addition, the variant is present in both sexes with the same manifestation. These findings are in contrast to other studies, what is very likely the result of previously mistaken variants. The study provides a detailed definition of Allen's fossa and a clear distinction from other variants at the femoral neck. The presented guidelines will allow a better comparability between different investigators in further studies. Individual differences, including vascular deficits and pathological conditions, can cause a considerable amount of variations in the manifestation of Allen's fossa, which is most likely caused by grinding of the zona orbicularis on the femoral neck.

Profiling of human burned bones: oxidising versus reducing conditions

Complementary optical and neutron-based vibrational spectroscopy techniques (Infrared, Raman and inelastic neutron scattering) were applied to the study of human bones (femur and humerus) burned simultaneously under either aerobic or anaerobic conditions, in a wide range of temperatures (400 to 1000 °C). This is the first INS study of human skeletal remains heated in an oxygen-deprived atmosphere. Clear differences were observed between both types of samples, namely the absence of hydroxyapatite’s OH vibrational bands in bone burned anaerobically (in unsealed containers), coupled to the presence of cyanamide (NCNH2) and portlandite (Ca(OH)2) in these reductive conditions. These results are expected to allow a better understanding of the heat effect on bone´s constituents in distinct environmental settings, thus contributing for an accurate characterisation of both forensic and archaeological human skeletal remains found in distinct scenarios regarding oxygen availability.

Source of strontium in archaeological mobility studies\u2014marine diet contribution to the isotopic composition

The strontium isotope composition of human tissues is widely used in archaeological mobility studies. However, little attention is paid to the relative contributions of terrestrial versus marine sources of strontium in these studies. There is some debate over the role of a solid diet versus drinking water as the most important source of strontium for the human body, with related possibilities of misinterpretation of the archaeological record if only strontium isotope compositions of the biosphere are studied. However, there is a third component, marine strontium, which is commonly not assumed to contribute towards the strontium isotope composition of archaeological skeletal remains, especially in locations that are not directly coastal. To illustrate the potentially obfuscating effects of mixed Sr sources in a human population, we present a case study of twelve individuals from the medieval Finnish site Iin Hamina with a known dietary history. Our study shows that marine consumption is a significant factor explaining the strontium isotope composition of the Iin Hamina human remains, with implication of erroneous conclusions about immigration without prior knowledge of diet composition. Thus, future studies should always incorporate a rigorous analysis of dietary history, with special regard to potential consumption of aquatic resources, when strontium isotope analysis is used as a method in the study of palaeomobility.

Comparing biological and pathological factors affecting osteocalcin concentrations in archaeological skeletal remains

Paleoproteomics and the study of ancient proteins has become an important consideration in bioarchaeological research as we seek to understand the relationship between the physical skeleton and its underlying biochemistry. Osteocalcin is an abundant, non-collagenous protein that is accessible archaeologically due to its affinity for hydroxyapatite in bone. Manufactured by the osteoblast cells during normal bone metabolism, osteocalcin is inherently linked to the process of bone remodelling. Clinical studies have consistently shown the impact of biological (i.e., sex and age) and pathological (i.e., disease) factors on osteocalcin concentrations within the body; however, this connection has yet to be fully explored bioarchaeologically. For this study, 46 individuals from the Danish Sct. Albert’s cemetery (CE 1250–1536) were analyzed with cortical bone samples harvested from the femur. Each sample was demineralized and quantified using established paleoproteomic methods (ELISA) and tested for diagenesis using FTIR-ATR. No significant differences were found in the osteocalcin concentrations between adult males and females. However, males had a wider range of osteocalcin variability perhaps explained in part by their increased prevalence of pathological conditions characterized by bone remodelling. Similarly, non-adults had a wide range of osteocalcin variability but there were no significant differences when compared to the adult mean. No correlations were found between osteocalcin concentrations and diagenesis suggesting that the protein can survive across a range of temporal periods and variable burial environments. While no clear pattern emerged between osteocalcin concentrations and biological and/or pathological factors, this study is significant in that it further explores the use of paleoproteomics in bioarchaeological contexts and how we may unite our macroscopic and biochemical assessments of human skeletal remains.

Paleogenomic insights into the red complex bacteria Tannerella forsythia in Pre-Hispanic and Colonial individuals from Mexico.

The 'red complex' is an aggregate of three oral bacteria (Tannerella forsythia, Porphyromonas gingivalis and Treponema denticola) responsible for severe clinical manifestation of periodontal disease. Here, we report the first direct evidence of ancient T.forsythia DNA in dentin and dental calculus samples from archaeological skeletal remains that span from the Pre-Hispanic to the Colonial period in Mexico. We recovered twelve partial ancient T. forsythia genomes and observed a distinct phylogenetic placement of samples, suggesting that the strains present in Pre-Hispanic individuals likely arrived with the first human migrations to the Americas and that new strains were introduced with the arrival of European and African populations in the sixteenth century. We also identified instances of the differential presence of genes between periods in the T. forsythia ancient genomes, with certain genes present in Pre-Hispanic individuals and absent in Colonial individuals, and vice versa. This study highlights the potential for studying ancient T. forsythia genomes to unveil past social interactions through analysis of disease transmission. Our results illustrate the long-standing relationship between this oral pathogen and its human host, while also unveiling key evidence to understand its evolutionary history in Pre-Hispanic and Colonial Mexico. This article is part of the theme issue 'Insights into health and disease from ancient biomolecules'.

Assessing the reliability of virtual reconstruction of mandibles

Mandibular morphological variation is often used to examine various aspects of human palaeobiology. However, fossil and archeological skeletal remains are often fragmented/distorted and so are frequently excluded from studies. This leads to decreased sample sizes and, potentially, to biased results. Thus, it is of interest to restore the original anatomy of incomplete/distorted specimens. Thin plate splines (TPS), commonly used in Geometric Morphometrics (GM), offer the prospect of reconstruction of missing parts and particularly of interest here, missing landmarks. Here, the reliability of TPS based mandibular reconstruction is tested. To that end missing landmarks were simulated in originally complete hemimandibles. TPS was then used to restore the location of simulated missing data and the predicted landmarks were compared to the original ones. Results show that error varies according to the number and location of estimated landmarks. Notwithstanding, estimation error is usually considerably smaller than the morphological differences between individuals from the same species. TPS based reconstruction allows fragmentary mandibles to be used in studies of whole mandibular variation, provided the above mentioned caveats are considered.

Palaeoproteomic identification of breast milk protein residues from the archaeological skeletal remains of a neonatal dog

Accurate postmortem estimation of breastfeeding status for archaeological or forensic neonatal remains is difficult. Confident identification of milk-specific proteins associated with these remains would provide direct evidence of breast milk consumption. We used liquid chromatography coupled to tandem mass spectrometry (MS) to confidently identify beta-lactoglobulin-1 (LGB1) and whey acidic protein (WAP), major whey proteins associated with a neonatal dog (Canis lupus familiaris) skeleton (430–960 cal AD), from an archaeological site in Hokkaido, Japan. The age at death of the individual was estimated to be approximately two weeks after birth. Protein residues extracted from rib and vertebra fragments were analyzed and identified by matching tandem MS spectra against the dog reference proteome. A total of 200 dog protein groups were detected and at least one peptide from canine LGB1 and two peptides from canine WAP were confidently identified. These milk proteins most probably originated from the mother’s breast milk, ingested by the neonate just before it died. We suggest the milk diffused outside the digestive apparatus during decomposition, and, by being absorbed into the bones, it partially preserved. The result of this study suggests that proteomic analysis can be used for postmortem reconstruction of the breastfeeding status at the time of death of neonatal mammalian, by analyzing their skeletal archaeological remains. This method is also applicable to forensic and wildlife studies.

Leukemia in Ancient Egypt: Earliest case and state‐of‐the‐art techniques for diagnosing generalized osteolytic lesions

AbstractMalignant bone tumors are uncommon among archaeological skeletal remains, and the general skeletal involvement is even less frequent. Multiple myelomas along with metastases are the most common conditions in paleopathology, whereas malignant myeloproliferative diseases have only been described in a handful of cases.We present a probable case of acute lymphocytic leukemia in a skeleton of an individual from Ancient Egypt, dated to the end of the First Intermediate Period and the beginning of the Middle Kingdom (c 2160–2000 bc). The excavation of the Temple of Millions of Years of Thutmosis III located in el‐Assasif (Luxor, Upper Egypt) recovered a total of 41 complete individual from an associated grave‐Necropolis placed close to the north enclosure wall. The individual, a 16 to 21‐year‐old male, showed a severe disease affecting all the skeleton with, predominantly, osteolytic lesions and areas of new bone formation. Gross examination under magnification and X‐ray in field were performed.We review the paleopathological literature to compare the cases previously reported of malignant disease. The morphology and distribution of the lesions of the individual are discussed to make an adequate differential diagnosis in the individual. Several criteria suggest that the disease was most likely acute lymphocytic leukemia: The individual died before the age of 21, the patterns of the bone lesions as well as its distribution below the elbow and knee joints, the presence of a gap in the coronal suture, and the mandible involvement.If this is the case, it would be the oldest case of leukemia described in Ancient Egypt and could help to understand the evolution patterns of cancer in the history.

Seaweed fertilisation impacts the chemical and isotopic composition of barley: Implications for analyses of archaeological skeletal remains

Fertilisation with animal manure has been shown to affect crop chemical and isotopic composition, indicating that if manuring effects are not taken into account, there is a risk of overestimating consumer trophic levels in palaeodietary studies. The effect of fertilisation with seaweed, a common fertiliser in the past in coastal areas, has been the subject of several hypotheses, but until now has not been studied in this particular context.In this study the impact of fertilising bere, an ancient type of Scottish barley (Hordeum vulgare L.), with 25 t/ha and 50 t/ha seaweed, in comparison to a modern commercial mineral fertiliser and to no fertilisation, was investigated in a field trial on the Orkney Islands, Scotland. Stable isotope ratios (δ13C and δ15N) and elemental concentrations (B, Mg, K, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Sr, Mo, Cd and Pb) of grain, husk and straw samples were determined. Significant differences were found between treatment groups, including increases in δ15N values of 0.6 ± 0.5‰ (average ± 1σ for five replicate plots) in grain, and 1.1 ± 0.4‰ in straw due to seaweed fertilisation. Elevated concentrations of Sr in grain and husk samples (factors of 1.2–1.4) indicate the geographic tracer 87Sr/86Sr may also be affected.Fertilisation with seaweed thus needs to be considered for archaeological interpretations of chemical and isotopic compositions of crop and skeletal material for accurate palaeodietary and provenance reconstructions, particularly in coastal areas. Further implications of these results for studies concerning the effects of sea spray, radiocarbon-dating, and for dietary reconstructions using trace elements are also identified.