Human Skeletal Remains Research Articles

From Jane Doe to Sofia: DNA Extraction Protocol from Bones and Teeth without Liquid Nitrogen for Identifying Skeletal Remains.

DNA analysis plays a crucial role in forensic investigations, helping in criminal cases, missing persons inquiries, and archaeological research. This study focuses on the DNA concentration in different skeletal elements to improve human identification efforts. Ten cases of unidentified skeletal remains brought to the Institute of Forensic Medicine in Timisoara, Romania, underwent DNA analysis between 2019 and 2023. The results showed that teeth are the best source for DNA extraction as they contain the highest concentration of genetic material, at 3.68 ng/µL, compared to the petrous temporal bone (0.936 ng/µL) and femur bone (0.633 ng/µL). These findings highlight the significance of teeth in forensic contexts due to their abundant genetic material. Combining anthropological examination with DNA analysis enhances the understanding and precision of identifying human skeletal remains, thus advancing forensic science. Selecting specific skeletal elements, such as the cochlea or teeth, emerges as crucial for reliable genetic analyses, emphasizing the importance of careful consideration in forensic identification procedures. Our study concludes that automated DNA extraction protocols without liquid nitrogen represent a significant advancement in DNA extraction technology, providing a faster, more efficient, and less labor-intensive method for extracting high-quality DNA from damaged bone and tooth samples.

Examining evidence for violence at the end of the Assyrian trading colonies period at Kaman‐Kalehöyük

AbstractThe end of the Assyrian Trading Colonies Period (ATCP) in Anatolia represents a time of significant change in the region. Historical and archaeological evidence suggests that considerable conflict and turmoil were associated with the end of the ATCP, but thus far little evidence from human skeletal remains has been available to support this hypothesis. One of the largest human skeletal samples dating to the end of the ATCP has been excavated at Kaman‐Kalehöyük in central Anatolia. A minimum of 73 individuals have been found in and around areas of the site that have signs of destruction and burning. These individuals appear to be associated with a battle that occurred in the town in the mid‐18th century BCE. The results show that the victims of the battle included individuals of all age groups and both sexes. The degree of burning and fragmentation made the identification of perimortem trauma challenging; however, perimortem trauma was observed in at least two individuals. An additional five individuals had evidence of antemortem trauma that was likely the result of interpersonal violence prior to the attack on the town. The skeletal evidence for trauma further supports the idea that a battle occurred at the site. This battle may have also been a part of a regional pattern of violence corresponding with the end of the ATCP, as suggested by the textual and archaeological evidence.

Unveiling intra-skeletal variability in mature and immature human skeletal remains via ATR-FTIR spectroscopy

Molecular characteristics of skeletal remains were studied utilizing ATR-FTIR spectroscopy, focusing on comparisons between mature adult and immature non-adult skeletal elements. To cover the intra-skeletal variability, different types of bones were analysed. The objective was to identify significant differences between various skeletal elements of adults and non-adults. Additionally, the correlation between observed differences and DNA preservation was investigated. Despite exposure to taphonomic factors, findings indicate minimal diagenetic changes or a well-balanced alteration in mineral and collagen within bones. The identified differences primarily reflect functional and structural differences among various skeletal elements. Significant differences between adults and non-adults, or lack of it, is attributed to different paths of bone maturation from childhood to adulthood. High DNA preservation in non-adult petrous bones was attributed to the interplay between DNA and carbonates, both occupying hydroxyl sites in the lattice. Conversely, lower DNA content in other bones, especially non-adult bones, was correlated with high relative concentrations of collagen, in which DNA is less stable and more prone to degradation. This study highlights the importance of skeletal variation (inter, intra, developmental stage) when assessing the preservation state of the remains and choosing samples for further analyses such as DNA. For the first time, differences between mature adult and immature non-adult bones are included.

A decolonizing approach to digital 3D visualizations and 3D printed models of human skeletal remains as demonstrative evidence in court: a view from Aotearoa New Zealand

ABSTRACT This research examines the impact of creating and presenting 3D visualizations and 3D printed models of human skeletal remains in relation to the cultural and ethical values of Māori, the indigenous people of Aotearoa New Zealand. Within Aotearoa, there is a complex history where Māori have not been consulted regarding the introduction of new technologies and systems, which ultimately had negative consequences for Māori. Using a decolonizing research approach, interviews (N = 12) and a focus group (N = 2) were conducted to identify key concepts regarding the sovereignty of these novel forms of digital data through a Māori cultural framework. For Māori, human skeletal remains are sacred; this research explored key questions including Māori Data Sovereignty and Rights, and what the digital data from human remains represents for Māori. Particular themes were identified regarding consultation with whānau (extended family) about the usefulness of the data to justice outcomes, and the rights of whānau to access, storage and disposal of the digital data, and subsequent printed 3D models. The core discussion across all participants related to tikanga Māori (Māori protocols) regarding the creation and interpretation of data, and the need for wider consultation with forensic scientists, lawyers, judges, Police and iwi (tribes) across Aotearoa.

Technical note: Temperature estimation accuracy based on colourimetry of embalmed human and fresh non-human burned bone

Estimation of the exposure temperature of skeletal remains can be done by means of colourimetry and a previously published decision model for the colourimetric data, resulting in clusters that represent a range of exposure temperature. The method was based on thermally altered freshly burned human skeletal remains. However, in practice the origin or pre-burning condition of (possibly fragmentary) burned remains can be unknown. Further, in order to use the colourimetric analysis, and accompanying decision model, it is important to have a reference or test set. Fresh human material is not available for this purpose in all countries. Hence, the classification accuracy of the decision model was tested for, in some countries more readily available substitutes for fresh bone; embalmed human bone and non-human bone. The model yielded high accuracies for these sample materials, making it possible to create a reference or test set from fresh non-human and embalmed human bone as substitute for human, and also to use the decision model for these deviating samples in practice.

Continuing Osteological Research with Fairmont State University’s Human Skeletal Collection

Human skeletal remains have been used at some point by most institutions involved in natural science education, though many real remains are slowly being replaced in classroom use by plastic models. Unfortunately, most antique teaching skeletons still in collections today were obtained during an ethically gray era, resulting in a lack of documentation and accountability. Since their purchase, many of these skeletal remains have been disassociated and damaged due to improper storage, the selling of partial skeletons, and mishandling. Projects such as this one at Fairmont State University are being performed to rehumanize and reassociate individuals using a variety of osteometric and macroscopic analysis techniques and create osteobiographies for them. Fairmont State University’s collection has been determined to contain a minimum of 43 individuals, 28 of whom were able to have profile information restored. Current ethical and legal standards related to the treatment of teaching skeletons are nearly non-existent in the US, but projects such as this one can bring attention to the historically immoral treatment of these individuals.

Association between morphological features in the cochlear promontory and mastoid Process: Implications for Identifying middle ear diseases in human skeletal remains

Infectious Middle Ear Disease (IMED) is a group of infections that frequently afflict individuals across all age groups worldwide. Although in today’s clinical practice IMED is efficiently managed, in the past it probably caused severe complications (e.g., hearing loss, balance problems, and facial nerve paralysis), heavily impacting the everyday life of many people. However, despite the valuable insights that studying ear infections in the past can bring, bioarcheology presently lacks a standardized methodology to assess their prevalence in skeletal populations. This can lead to several issues, including incomparability of results, difficulties in replications, and methodological biases.Recently, straightforward and accessible criteria for the observation of the cochlear promontory were presented to successfully assess IMED on skeletal remains. Yet, as otologists rarely examine the impact of ear infections on the bony structures of the middle ear, the results were not supported clinically, and could therefore not be validated. To fill this gap, we propose a study in which computed tomography (CT) of the mastoid process was utilized on the skulls of 50 individuals to verify the results obtained through gross observation of the cochlear promontory. Statistical analysis revealed significant correlation between hypopneumatization (indicative of childhood IMED) and bony changes of the promontory at the level of the individual. This suggests a potential correlation that warrants further investigation to determine whether the observation of the cochlear promontory should be used as a method for IMED assessment. Overall, our study contributes to the study of IMED in past populations, and underscores the importance of clear, standardized scoring criteria in paleopathology.

Zuurberg (Eastern Cape, South Africa) revisited: human remains, dating and archaeological findings

ABSTRACT The Later Stone Age (LSA) site of Zuurberg in the Cape Fold Mountains of South Africa was excavated in the 1920s by F.W. FitzSimons. As with his other excavations, not much is known about the exact provenance and detailed archaeological context of the remains. This paper reports the re-analysis and dating of the human skeletal remains from Zuurberg. The remains were commingled but here sorted according to previously published sources. The remains of eight individuals are present, of which only one (Zg3) is fairly complete. The assemblage includes four older adults, two younger or middle-aged adults, an adolescent and a child. Two of the three individuals who could be assessed showed signs of enamel hypoplasia, which probably attests to some hardship. One of the individuals was dated to 4795 ± 42 BP (Ua-61976), which places this site into the Wilton period of the South African LSA. However, the different levels of burials, as well as the presence of potsherds, may suggest an extended use of the site. Isotopic analysis suggests a mixed diet, which was not depleted of protein. The reassessment, sorting and dating of these remains add value to the collection, especially as they come from an open-air, non-coastal forager site. This assemblage can now be included in future studies of southern African Stone Age foragers.

Stature Estimation using Tibia Length in Young Adults of Urhobo Ethnic Group, in South-south, Nigeria

Introduction: Estimating stature is an essential part of the identification of human skeletal remains or body parts. The purpose of this study was to estimate stature using tibia length among young Urhobo Adults.
 Materials and method: 350 subjects (144 males and 206 females) were utilized for the study. Stature was measured using a stadiometer and tibia length was measured using a digital Vernier calliper. The independent and paired samples t-test, Pearson's correlation test and linear regression equations were employed using Statistical Package for Social Sciences (SPSS) software.
 Results: The mean height in males was 174.22cm and females was 163.97cm, the mean Right tibia length was 40.48cm in males and 37.67cm in females while the left tibia length was 40.41cm and 37.63cm in males and females respectively. There was significant difference between right and left tibia length in males and the general population (p<0.001). However, females showed no significant difference (p>0.05). There was a strong positive significant association between stature and bone lengths (r>0.8, p<0.001). There was no significant difference between the estimated and actual stature for subjects using the general and group-specific regression formulas (p>0.05).
 Conclusion: The study showed that males have significantly higher mean values of parameters measured than females. Right tibia length was significantly longer than the left in males and total population. Groupspecific formula gave more accurate results than the general formula making them better suited to estimate stature in forensic cases. Hence, tibia length can be used as an important tool in forensic examination as they provide elements of accuracy in stature determination.

X-chromosomal STRs in aDNA kinship analysis.

The analysis of ancient DNA (aDNA) from human skeletal remains can provide useful insights when investigating archaeological finds. One popular application of aDNA is to examine genealogical relationships between individuals recovered at the same archaeological site. For the reconstruction of genealogical relationships, several genetic markers are commonly used: autosomal STRs, mitochondrial lineages (based on SNP-analysis) and Y-chromosomal haplotypes (based on Y-STR-analysis). In this paper, we present the additional opportunities that X-STRs provide in aDNA kinship reconstruction, especially in deficiency cases and for the examination of father-daughter relationships. Possible applications are demonstrated on a range of different kinship reconstructions: confirmation of half-siblingship in the Lichtenstein cave (Germany), exclusion of two potential father-daughter relationships in Goslar (Germany), investigation of three siblingships in Boilstädt (Germany) as well as the confirmation of a father-daughter relationship in Stolpe (Germany). This study shows that the analysis of X-STRs can contribute to the investigation of relationship constellations otherwise difficult to approach (e.g. father-daughter relationships) and that X-STRs are useful to support and complement autosomal STRs, mtDNA and Y-STR data.

Solving cold cases: The importance of new identification techniques in old cases of forensic anthropology

Scientific and technological developments experienced by forensic sciences have increased the possibility of human identification. Forensic anthropology has developed worldwide and has made significant progress in Brazil in the last 10 years. This context has allowed forensic medicine institutes to use new techniques to resolve cold cases of human identification. This work aimed to report the methodology, development, and partial results of the LAFOL Identifies Operation, carried out at the Forensic Anthropology and Forensic Dentistry Laboratory (LAFOL) of the Medico-Legal Institute of Porto Velho. We carried out a multidisciplinary expert approach on human skeletal remains archived over the last 20 years to identify these individuals using current scientific methods. Eighty-one cold cases of skeletonized and/or mummified human skeletal remains were reanalyzed; they were archived in the LAFOL Laboratory collection as unidentified. Anthropological analyses showed that the collection is predominantly composed of adult individuals (86.4%), between 21 and 39 years old (55.6%), and males (62.9%). In the human skeletal remains identified, forensic anthropology played a key role in the case screening process. We expected that the results presented in this work will inspire other Medico-Legal Institutes in Brazil and worldwide to establish task forces for the reanalysis of cold cases in their institutes, promoting new forensic dentistry and anthropological comparisons and collecting biological samples to feed the database of genetic profiles of missing persons.

Intensified exploitation of animal products in the Mediterranean Copper Age: isotopic evidence from Scaba ’e Arriu (Sardinia)

In this paper, dietary patterns are reconstructed across two phases represented at the Copper Age of Scaba ’e Arriu (Sardinia, Italy) via isotopic analysis of human and animal skeletal remains. Collagen carbon, nitrogen, and bioapatite carbon, and oxygen isotopes were used to infer diet, economic practices, and possibly different climatic conditions. Differential management of cattle, sheep/goats, and pigs was detected, with scarce animal products in the human diet in the Early Copper Age, followed by an increase in animal products identifiable in herbivore livestock, coincident with more rainy conditions in the Late Copper Age. Dietary information was then considered in light of the climatic periods already known for the period and compatible with the data presented. The study’s outcome indicates generally sedentary and endogamous groups with only a few outliers in both phases.

Biting into the truth: Connecting oral pathology and stable isotopes through the paradigmatic example of a hyper-specialized marine diet in Medieval Pontevedra (NW Iberia)

Human feeding patterns have been reconstructed in Archaeology by analysing either oral pathology or stable isotope ratios in human skeletal remains. However, no clear agreement has been developed between these two methodologies. The main objective of this study is to determine if we can establish a link between them when analysing a population with a hyper-specialized diet, in this case marine resources (and millet/maize). To reach this goal we developed a conjoined multi-isotope analysis using collagen and bioapatite (δ13Ccol, δ15Ncol and δ13Ccar) and a detailed study of oral health (caries, antemortem loss, periapical lesions, periodontal disease, calculus, and wear). All available skeletons with at least one preserved tooth from two cemeteries of the medieval town of Pontevedra (n = 34) were studied. The buried individuals belonged to the guild of fishers and artisans, professions which were dominant among the families of medieval Pontevedra. A detailed FTIR-ATR study of extracted bone bioapatite showed a high correlation between bioapatite carbonate content, carbonate typical vibrations, and FTIR-ATR indices related to bone diagenesis, which is in line with previous research. No significant correlations were found with bioapatite yield and isotopic composition (δ13Ccar and Δ13C), ruling out possible diagenetic effects. The diet was based on marine fish protein with contributions of millets (e.g., δ13Ccar -11.9 ± 1.8‰) that seems to be slightly higher in individuals linked to artisanal guilds. The oral pathology study shows severe dental wear from an early age (Grade 2–4 in permanent dentition for 20% of infants and 60% of juveniles in M1), as well as moderate-high presence of caries in permanent dentition (64%, 22/34) and dental calculus (72%, 24/33). Both the oral pathology and the isotopic signal differ from that observed in other areas of the Iberian Peninsula. This study points to the existence of connections between findings of the two methodologies, and specifically an association between intense dental wear and high consumption of marine resources and millet. At the same time, this analysis implies the necessity of caution in estimation of age by dental wear in populations linked to the sea.

Periodontitis and alveolar resorption in human skeletal remains: The relationship between quantitative alveolar bone loss, occlusal wear, antemortem tooth loss, dental calculus and age at death in a low socioeconomic status, modern forensic human collection from Yucatan

ObjectiveThis paper assesses the relationship between the distance between the cemento-enamel junction and alveolar crest and risk factors commonly associated with periodontitis. MaterialsEighty individuals between 28 and 92 years old with known biological sex and age were analyzed from a 20th century forensic human collection from Merida, Yucatan (Mexico). MethodsMacroscopic assessment, along with metric analysis, was employed using a probe. ResultsAnte-mortem tooth loss was positively correlated with the distance between the cemento-enamel junction and alveolar crest, as was the presence of root calculus in females. ConclusionsCemento-enamel junction to alveolar crest distance is not a reliable indicator of periodontitis since it is not directly related to periodontitis-causing infectious pathogens, and since ante-mortem tooth loss can affect root exposure. SignificanceThis study demonstrates that a purely quantitative approach to diagnosing periodontitis in archaeological and forensic human remains can be misleading. LimitationsThe skeletal collection is only representative of the low socioeconomic class of Merida, and its female cohort is underrepresented. In addition, because the Xoclan collection is modern, limitations (particularly with respect to tooth wear) of the applicability of these interpretations to older archaeological remains exist. Suggestion for further researchA combination of quantitative and qualitative characteristics of alveolar bone is needed to reliably diagnose periodontitis in skeletal populations.

Sex differences in mechanical stress and associated labor in Hellenistic–early Roman Menainon, Sicily

AbstractThis paper explores mechanical stress in the Hellenistic/early Roman community of Menainon (Sicily) to test the existence of sex‐based division of labor. For this purpose, human skeletal remains from 79 males and 65 females, dating to the 4th–1st ca. BCE, were examined for degenerative joint disease, intervertebral disc disease, cross‐sectional geometric properties, and entheseal changes. Our findings support an equal share of workload between males and females, who appeared to have experienced similar mechanical stress levels. In contrast, they do not support a systematically greater mechanical load among males, as would have been expected if females had been restricted indoors taking care of domestic tasks only, while men had dealt with most physically demanding outdoor activities. The patterns identified in the current study agree with a scenario where agriculture was a prominent subsistence activity (as attested historically for this region) and most members of this rural community engaged in agricultural tasks irrespective of sex. Our study has limitations regarding the available historical and archaeological information, which restrict our ability to formulate solid research hypotheses or fully contextualize the bioarchaeological results. Nonetheless, it does highlight the importance of critically assessing historical information regarding the role of men and women in the Greco‐Roman society.

Life and death in the Roman period Pula – A bioanthropological analysis of human skeletal remains from the Ozad Arene site

Human skeletal and dental remains are the primary focus of bioarchaeological research, as different diseases and pathological conditions can leave marks on the bones and teeth. Study of these remains provides insights into the lifestyle, health and quality of life of past populations. In this paper, the human skeletal remains from the Ozad Arene archaeological site in Pula (Istria, Croatia), originating from 25 graves and four bone assemblages dated between the 1st and the 4th centuries CE, were analysed. The aim of the paper is to reconstruct demographic and pathological characteristics of the Roman period population from Pula to gain insight into the living conditions and the quality of life in this community, and to determine whether the site differs from other archaeological sites from the same period in terms of general health and living conditions. The human skeletal sample in question comprises skeletal remains of a minimum 48 individuals; the high subadult mortality rate and high prevalence of dental enamel hypoplasia and the presence of cribra orbitalia, porotic hyperostosis, and periostitis indicate overall poor living conditions and widespread metabolic stress during childhood in Pula. Vertebral osteoarthritis and Schmorl′s nodes in the spines of younger individuals suggest continuous hard labour and strenuous physical activity. Based on the available data, it seems that living conditions in the Roman period Pula did not differ significantly from other Roman sites on the eastern Adriatic coast.

Ballistic long bone fracture pattern: an experimental study.

When dealing with badly preserved cadavers or skeletal human remains, the assessment of death circumstances remains challenging. When forensic evidence cannot be taken from the skin and soft tissue, the information may only be deduced from more resistant elements such as bone. Compared to cranial gunshot injuries, reliable data on ballistic long bone trauma remains scarce. This study aims to define ballistic fracture characteristics in human long bones. The shaft of 16 femurs and 13 humeri from body donors was perpendicularly shot with a 9-mm Luger full metal jacket bullet at an impact velocity of 360m/s from a distance of 2m. Some bones were embedded in Clear Ballistics Gel®, and some were shot without soft tissue simulant in order to better visualise the fracture propagation on the high-speed camera. The fractures were examined macroscopically and compared between the sample groups. We consistently found comminuted fractures with a stellate pattern. Fracture details were classified into entrance, exit and general characteristics. For some traits, we detected different occurrence values in the group comparison. The results indicate that some of the traits depend on bone properties such as shaft diameter, bone length and cortical thickness. The presence of ballistic gel also influenced some fracture traits, emphasising the relevance of soft tissue simulant in osseous gunshot experiments. This study revealed new insights in the detailed fracture pattern of human long bones. These may serve as guidelines for the identification and reconstruction of gunshot trauma in human long bones.

Application of X‐ray fluorescence in forensic archeology: A review

AbstractForensic Archeology is the application of techniques and principles of archeology in the pursuit of examining a crime or incident with legal interest. A wide range of analytical studies are employed in examining archeological remains. X‐ray fluorescence (XRF) spectroscopy remains to be one of the most used techniques for rapid examination of trace evidence typically found on an archeological crime scene. Studies of interest generally include finding trace elements from various sources, samples, and different environmental conditions and distinguishing whether a skeletal remains under examination is human, animal, or marine species. The present study has been designed to focus on the application of XRF in archeological and anthropological evidences such as the examination of human skeletal and dental remains, determination of species, examination of soil, ceramics, paintings, coins, and so on. The advantage of XRF as derived from the reported literature is that it is a non‐destructive technique making it ideal for forensic analysis and in situ examination. The study also discusses the factors affecting forensic investigations of archeological evidences and the limitations of XRF.

Bioanthropological analysis of human remains from the archaic and classic period discovered in Puyil cave, Mexico.

Determine the geographic place of origin and maternal lineage of prehistoric human skeletal remains discovered in Puyil Cave, Tabasco State, Mexico, located in a region currently populated by Olmec, Zoque and Maya populations. All specimens were radiocarbon (14C) dated (beta analytic), had dental modifications classified, and had an analysis of 13 homologous reference points conducted to evaluate artificial cranial deformation (ACD). Following DNA purification, hypervariable region I (HVR-1) of the mitogenome was amplified and Sanger sequenced. Finally, Next Generation Sequencing (NGS) was performed for total DNA. Mitochondrial DNA (mtDNA) variants and haplogroups were determined using BioEdit 7.2 and IGV software and confirmed with MITOMASTER and WebHome softwares. Radiocarbon dating (14C) demonstrated that the inhabitants of Puyil Cave lived during the Archaic and Classic Periods and displayed tabular oblique and tabular mimetic ACD. These pre-Hispanic remains exhibited five mtDNA lineages: A, A2, C1, C1c and D4. Network analysis revealed a close genetic affinity between pre-Hispanic Puyil Cave inhabitants and contemporary Maya subpopulations from Mexico and Guatemala, as well as individuals from Bolivia, Brazil, the Dominican Republic, and China. Our results elucidate the dispersal of pre-Hispanic Olmec and Maya ancestors and suggest that ACD practices are closely related to Olmec and Maya practices. Additionally, we conclude that ACD has likely been practiced in the region since the Middle-Archaic Period.

Handheld hyperspectral imaging as a tool for the post-mortem interval estimation of human skeletal remains

In forensic medicine, estimating human skeletal remains' post-mortem interval (PMI) can be challenging. Following death, bones undergo a series of chemical and physical transformations due to their interactions with the surrounding environment. Post-mortem changes have been assessed using various methods, but estimating the PMI of skeletal remains could still be improved. We propose a new methodology with handheld hyperspectral imaging (HSI) system based on the first results from 104 human skeletal remains with PMIs ranging between 1 day and 2000 years. To differentiate between forensic and archaeological bone material, the Convolutional Neural Network analyzed 65.000 distinct diagnostic spectra: the classification accuracy was 0.58, 0.62, 0.73, 0.81, and 0.98 for PMIs of 0 week–2 weeks, 2 weeks–6 months, 6 months–1 year, 1 year–10 years, and >100 years, respectively. In conclusion, HSI can be used in forensic medicine to distinguish bone materials >100 years old from those <10 years old with an accuracy of 98%. The model has adequate predictive performance, and handheld HSI could serve as a novel approach to objectively and accurately determine the PMI of human skeletal remains.