What we see is what we touch? Sex estimation on the skull in virtual anthropology.

BACKGROUND: The increased use of virtual bone images in forensic anthropology requires a comprehensive study on the observational errors between dry bones and CT reconstructions. Here, we focus on the consistency of nonmetric sex estimation traits on the human skull. MATERIALS AND METHODS: We scored nine nonmetric traits on dry crania and mandibles (n = 223) of archaeological origin and their CT reconstructions. Additionally, we 3D surface scanned a subsample (n = 50) and repeated our observations. Due to the intricate anatomy of the mental eminence, we split it into two separate traits: the bilateral mental tubercles and the midsagittal mental protuberance. We provide illustrations and descriptions for both these traits. RESULTS: We obtained supreme consistency values between the CT and 3D surface modalities. The most consistent cranial traits were the glabella and the supraorbital margin, followed by the nuchal crest, zygomatic extension, mental tubercles, mental protuberance, mental eminence, mastoid process and ramus flexure, in descending order. The mental tubercles show higher consistency scores than the mental eminence and the mental protuberance. DISCUSSION: The increased interchangeability of the virtual modalities with each other as compared to the dry bone modality could be due to the lack of tactility on both the CT and surface scans. Moreover, tactility appears less essential with experience than a precise trait description. Future studies could revolve around the most consistent cranial traits, combining them with pelvic traits from a previous study, to test for accuracy.

What we see is what we touch? Sex estimation on the pelvis in virtual anthropology.

BACKGROUND: Computed tomography (CT) scans are a convenient means to study 3D reconstructions of bones. However, errors associated with the different nature of the observation, e.g. visual and tactile (on dry bone) versus visual only (on a screen) have not been thoroughly investigated. MATERIALS AND METHODS: We quantified the errors between modalities for sex estimation protocols of nonmetric (categorical and ordinal) and metric data, using 200 dry pelves of archaeological origin and the CT reconstructions of the same bones. In addition, we 3D surface scanned a subsample of 39 pelves to compare observations with dry bone and CT data. We did not focus on the sex estimation accuracy but solely on the consistency of the scoring, hence, the interchangeability of the modalities. RESULTS: Metric data yielded the most consistent results. Among the nonmetric protocols, ordinal data performed better than categorical data. We applied a slightly modified description for the trait with the highest errors and grouped the traits according to consistency and availability in good, intermediate, and poor. DISCUSSION: The investigated modalities were interchangeable as long as the trait definition was not arbitrary. Dry bone (gold standard) performed well, and CT and 3D surface scans performed better. We recommend researchers test their affinity for using virtual modalities. Future studies could use our consistency analysis and combine the best traits, validating their accuracy on various modalities.

Quantification of dental macrowear using 3D occlusal surface topographic measurements in deciduous and permanent molars of children.

OBJECTIVES: Childhood paleodietary reconstruction via dental macrowear analysis is limited in part by available methods to measure dental macrowear. We describe a method to quantify dental macrowear progression (in both deciduous and permanent molars) using a handheld intraoral scanner and two 3D occlusal topographic measurements. We assess the agreement of our macrowear proxies with an established qualitative wear scoring system and their relationship to age. MATERIAL AND METHODS: We scanned 92 well-preserved dentitions of immature individuals from the medieval cemetery of St. Pölten in Lower Austria using an intraoral scanner. Two measurements were made on the resulting mesh files-the relative flat surface area in % of the occlusal surface (RFSA%) and the mesial interior slope angle. We estimated the technical error of measurement (TEM). Comparisons were made with the macrowear scoring system-tooth wear index. RESULTS: We found that TEM for both measurements was between 1 and 3%, except the interobserver TEM of RFSA% which was above 5%. Both quantitative measurements generally agree with the established qualitative scores and correlate with age; however, RFSA% does not reliably indicate the progression of macrowear for teeth after dentine exposure occurs. DISCUSSION: The proposed 3D topographic measurements can be made reliably, and within a certain range of wear provide good quantitative proxies of the progression of dental macrowear. Such measurements constitute a promising approach for improving dental macrowear analysis in contexts such as childhood paleodietary reconstruction, which benefit from additional precision in wear rate estimation and present less dentine exposure.

Death due to coronary artery anomaly with coexistence of Chiari network.

Coronary artery anomalies rarely detected in autopsy series and angiograms can be a component of complex malformations, besides, can be also associated with sudden cardiac death. Presented case was 22-year-old male, who had suddenly fainted during a football match played on artificial turf, he was transferred into the hospital, however had died during intensive care therapy. He had been evaluated by local prosecutor, and sent to our center for autopsy. At autopsy, internal macroscopic examination revealed absence of the right coronary artery. A total of two coronary artery ostia were observed. One of them originated from the left aortic sinus, and the other one stemmed from 8 mm above the sinotubular line. Besides, Chiari network formation was seen in the right atrium. This case with coronary artery anomaly associated with formation of Chiari network was discussed from the perspective of forensic medicine in the light of the literature information.

More than just a color: Archaeological, analytical, and procedural aspects of Late Bronze Age purple-dye production at Cape Kolonna, Aegina.

Excavations in the Eastern Suburb of Bronze Age Aegina Kolonna revealed the destruction deposit of two sequenced Early Mycenaean buildings (phase Late Helladic IIA; 16th century BC). The older building is interpreted as a widely undisturbed production site of purple-dye based on indicative finds such as ceramic sherds containing analyzable quantities of pigment, high amounts of mollusk shells, and a few functional facilities. Chemical analysis by HPLC and malacological determination revealed that the banded dye-murex (Hexaplex trunculus) was used almost exclusively. The presence of crushing tools and a waste disposal pit provide insight into the technical process of dye production. Additionally, skeletal remains of heavily burnt infantile and juvenile piglets, kids, or lambs were found in the purple workshop area. The evidence may be better explained by ritual activities aimed at promoting the highly meaningful event of purple production, rather than by normal food consumption practices.

Dental wear and oral pathology among sex determined Early Bronze-Age children from Franzhausen I, Lower Austria.

The physical properties of diet and oral health throughout childhood play an important role in the development of human dentition, and differed greatly before the industrial revolution. In this study we examined dental wear and oral pathology in a sample of children from the Early Bronze-Age to investigate the physical and mechanical properties of childhood diet and related oral health. We explore cross-sectional age and sex-based variation of children in the sample. The analysis was carried out on the dentitions of 75 children, 978 teeth, excavated from the Early Bronze-Age cemetery Franzhausen I in Lower Austria. Presence of dental caries and calculus was recorded. Dental wear was measured using dentine exposure, occlusal topography, and dental microwear texture analysis. Sex determination was carried out using amelogenin peptide analysis. Caries were found in only 4 individuals (crude prevalence rate-5%, 95% CI 1% to 13%), affecting only 5 teeth (true prevalence rate-less than 1%). Dentine exposure was observed in over 70% of deciduous molars and dental wear measurements indicate a comparatively strong dental wear accumulation especially, among younger children, when compared to modern-day and later pre-industrial populations. Microwear textures presented a high complexity (Asfc > 2)/low anisotropy (epLsar < 1) profile, especially in older children. Differences between male and female children were not generally significant but increased dentine exposure was observed in the lower molars of younger female children. Our results suggest that the Early Bronze-Age children at Franzhausen I consumed a non-cariogenic diet, more abrasive and inclusive of harder/polyhedral foodstuffs than present-day children and some later Medieval children. Differences in dental wear accumulation were observed between children within the population, but with minimal variation between the sexes mostly occurring among younger children.

Amelogenin peptide analyses reveal female leadership in Copper Age Iberia (c. 2900-2650 BC).

Given the absence of written records, the main source of information available to analyze gender inequalities in early complex societies is the human body itself. And yet, for decades, archaeologists have struggled with the sex estimation of poorly preserved human remains. Here we present an exceptional case study that shows how ground-breaking new scientific methods may address this problem. Through the analysis of sexually dimorphic amelogenin peptides in tooth enamel, we establish that the most socially prominent person of the Iberian Copper Age (c. 3200-2200 BC) was not male, as previously thought, but female. The analysis of this woman, discovered in 2008 at Valencina, Spain, reveals that she was a leading social figure at a time where no male attained a remotely comparable social position. Only other women buried a short time after in the Montelirio tholos, part of the same burial area, appear to have enjoyed a similarly high social position. Our results invite to reconsider established interpretations about the political role of women at the onset of early social complexity, and question traditionally held views of the past. Furthermore, this study anticipates the changes that newly developed scientific methods may bring to prehistoric archaeology and the study of human social evolution.

A case of congenital multiple epiphyseal dysplasia from the Late Migration Period graveyard in Drnholec (Czech Republic).

OBJECTIVE: To contribute to differential diagnosis of multiple epiphyseal dysplasia (MED) in archeological and clinical contexts. MATERIALS: A skeleton of a 30- to 45-year-old male (grave no. 806) from the Late Migration Period graveyard in Drnholec-Pod sýpkou (Czech Republic), radio-carbon dated to AD 492-530. METHODS: Morphological and metric analyses. RESULTS: Significant pathological changes were noted on ossa coxae and proximal ends of the femora, which appear similar to changes associated with Legg-Calvé-Perthes disease. X-ray examination made it possible to rule out pseudoachondroplasia, rickets and metabolic bone diseases. CONCLUSIONS: The finding was evaluated as a probable case of congenital multiple epiphyseal dysplasia. SIGNIFICANCE: This case will contribute to the construction of estimates of the occurrence of this disease in historical populations and can be instructive for diagnostics in current medical practice. LIMITATIONS: The final diagnosis is limited by the lack of genetic analysis. SUGGESTION FOR THE FUTURE RESEARCH: Further clarification leading to diagnosis will benefit from genetic analysis and evaluation of skeletal remains throughout Europe.

More than urns: A multi-method pipeline for analyzing cremation burials.

Burial rites of archaeological populations are frequently interpreted based on cremated remains of the human body and the urn they were deposited in. In comparison to inhumations, information about the deceased is much more limited and dependent on fragmentation, selection of body regions, taphonomic processes, and excavation techniques. So far, little attention has been paid to the context in which urns are buried. In this study, we combined archaeological techniques with anthropology, computed tomography, archaeobotany, zooarchaeology, geochemistry and isotopic approaches and conducted a detailed analysis on a case study of two Late Bronze Age urns from St. Pölten, Austria (c. 1430 and 1260 cal. BCE). The urns were recovered en-bloc and CT-scanned before the micro-excavation. Osteological and strontium isotope analysis revealed that the cremated remains comprised a young adult female and a child that died at the age of 10-12 years. Both individuals had been subject to physiological stress and were likely local. Animal bones burnt at different temperatures suggested different depositional pathways into the urn and pit as part of the pyre, food offerings, and unintentional settlement debris. Eight wild plant and five crop plant species appeared as part of the local landscape, as food offerings and fire accelerants. Sediment chemistry suggests that pyre remains were deposited around the urns during burial. Multi-element geochemistry, archaeobotany, and zooarchaeology provide insights into the Late Bronze Age environment, the process of cremation, the gathering of bones and final funerary deposition.

Common Ground between Biological Rhythms and Forensics.

Biological clocks set the timing for a large number of essential processes in the living human organism. After death, scientific evidence is required in forensic investigations in order to collect as much information as possible on the death circumstances and personal identifiers of the deceased victim. We summarize the associations between the molecular mechanisms of biological rhythms and forensically relevant aspects, including post-mortem interval and cause of death, entomological findings, sex, age, ethnicity and development. Given their importance during lifetime, biological rhythms could be potential tools to draw conclusions on the death circumstances and the identity of a deceased person by mechanistic investigations of the different biological clocks in a forensic context. This review puts the known effects of biological rhythms on the functions of the human organism in context with potential applications in forensic fields of interest, such as personal identification, entomology as well as the determination of the post-mortem interval and cause of death.

Single-incision cochlear implantation and hearing evaluation in piglets and minipigs.

OBJECTIVES: Various animal models have been established and applied in hearing research. In the exploration of novel cochlear implant developments, mainly rodents have been used. Despite their important contribution to the understanding of auditory function, translation of experimental observations from rodents to humans is limited due to the size differences and genetic variability. Large animal models with better representation of the human cochlea are sparse. For this reason, we evaluated domestic piglets and Aachen minipigs for the suitability as a cochlear implantation animal model with commercially available cochlear implants. METHODS: Four domestic piglets (two male and two female) and six Aachen minipigs were implanted with either MED-EL Flex24 or Flex20 cochlear implants respectively, after a step-by-step surgical approach was trained with pig cadavers. Electrophysiological measurements were performed before, during and after implantation for as long as 56 days after surgery. Auditory brainstem responses, electrocochleography as well as electrically and acoustically evoked compound action potentials were recorded. Selected cochleae were further analyzed histologically or with micro-CT imaging. RESULTS: A surgical approach was established using a retroauricular single incision. Baseline auditory thresholds were 27 ± 3 dB sound pressure level (SPL; auditory brainstem click responses, mean ± standard error of the mean) and ranged between 30 and 80 dB SPL in frequency-specific responses (0.5 - 32 kHz). Follow-up measurements revealed deafness within the first two weeks after surgery, but some animals partially recovered to a hearing threshold of 80 dB SPL in certain frequencies as well as in click responses. Electrically evoked compound action potential thresholds increased within the first week after surgery, which led to lower stimulation responses or increase of necessary charge input. Immune reactions and consecutive scalar fibrosis following implantation were confirmed with histological analysis of implanted cochleae and may result in increased impedances. A three-dimensional minipig micro-CT segmentation revealed cochlear volumetric data similar to human inner ear dimensions. CONCLUSIONS: This study underlines the feasibility of cochlear implantation with clinically used cochlear implants in a large animal model with representative inner ear dimensions comparable to humans. To bridge the gap between small animal models and humans in translational research and to account for the structural and size differences, we recommend the minipig as a valuable animal model for hearing research. First insights into the induced trauma in minipigs after cochlear implant surgery and a partial hearing recovery present important data of the cochlear health changes in large animal cochleae.

A histological comparison of non-human rib models suited for sharp force trauma analysis.

Cut marks provide essential knowledge to interpret which and how tools were used, both in archaeological and forensic context. Lots of studies focused on experimentally produced cut marks on animal models to develop methods for stabbing incidents. However, animal models are criticized to be morphologically different in comparison to human bones. This study analyzed the bone composition and experimentally obtained cut marks done on ribs from humans, pigs and goats. Methods included a qualitative description of differences between the species and a quantitative analysis of the cut mark proportions in histological thin sections and micro CT scans. The results indicated that especially the cortical bone of non-human ribs was different in comparison to human bone tissue as they were more robust and usually juvenile. Plexiform bone dominates and remodeled lamellar bone is rarely visible. The knife impact tends to create debris inside the cut mark and stress fractures along lamellae and cement lines perpendicular to the cut mark. Moreover, entheses of the intercostal muscles are always affected by the incision. Pig ribs were shown to be better suited as a model for sharp force trauma than goat ribs in comparison to humans. Though, plexiform bone and non-remodeled bone made it not quite ideal. We suggested to use bone material from animals used for breeding instead of meat production as they are slaughtered at higher ages.

Child murder in the Early Bronze Age: proteomic sex identification of a cold case from Schleinbach, Austria.

The identification of sex-specific peptides in human tooth enamel by nanoflow liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS) represents a quantum leap for the study of childhood and social relations more generally. Determining sex-related differences in prehistoric child rearing and mortality has been hampered by the insufficient accuracy in determining the biological sex of juveniles. We conducted mass spectrometric analysis to identify sex-specific peptides in the dental enamel of a child from a settlement pit of the Early Bronze Age settlement of Schleinbach, Austria (c. 1950-1850 bc). Four perimortal impression fractures on the skull of a 5-6-year-old child indicate an intentional killing, with a co-buried loom weight as possible murder weapon. Proteomic analysis, conducted for the first time on prehistoric teeth in Austria, determined the child's sex as male. While we cannot conclusively determine whether the child was the victim of conflicts between village groups or was slain by members of his own community, we suggest that contextual evidence points to the latter. A possible trigger of violence was the follow-on effects of an uncontrolled middle ear infection revealed by an osteological analysis. The boy from Schleinbach highlights the potential for further investigation of gender-biased violence, infanticide and child murder based on the recently developed method of proteomic sex identification.

Comparison of Bleaching Products With Up to 6% and With More Than 6% Hydrogen Peroxide: Whitening Efficacy Using BI and WI D and Side Effects - An in vitro Study.

The aim of this study was to evaluate the effect of bleaching agents containing different concentrations of hydrogen peroxide (HP) on color-change and on enamel-surface in bovine teeth. Furthermore the influence on cell viability and proliferation was investigated. Two hundred and forty teeth were randomly assigned into four groups (home bleaching ≤6%, in-office bleaching ≤6%, in-office bleaching > 6% HP, and control group). Bleaching was performed after artificial staining and the bleached index (BI) as well as the whiteness index (WI D ) was measured at several time points. Chemical analysis for HP concentrations and the pH of the bleaching products was done. Furthermore, enamel surfaces of randomly selected specimens were analyzed using scanning electron microscopy (SEM) and cytotoxicity of the tested bleaching products was evaluated in vitro using dental pulp cells (DPCs) and L929 cells. A statistically significant whitening effect was observed in almost all products. As expected all investigated products resulted in decreased cell viability, however, with different values of LC50 (median lethal concentration). SEM analysis showed an analog of enamel alterations with decreasing pH, increasing exposure time, and increasing HP concentration. Bleaching agents containing a low HP concentration are considered to be effective and to have less damaging effects on enamel and tested cells.

Head injuries of Roman gladiators.

Gladiator remains from a recently unearthed cemetery in ancient Ephesus (Turkey) offer a unique opportunity for proving common theories involving the weaponry and techniques of gladiator fighting based on the evidence supplied by cranial bones. This mass grave is the first of its kind to undergo a thorough osteological and forensic examination. A minimum number of individuals (MNI) analyses revealed that at least 68 individuals. All individuals found turned out to have been males aged between 20 and 30 years, except for one female associated with a female slave gravestone, and one male aged 45-55 years, had been buried in this area of the cemetery. The male mean body height was 168 cm (S.D.=5 cm), which lies inside the normal range of height for Roman populations at those times. Eleven (16% of MNI) individuals exhibit a total of 16 well-healed antemortal cranial traumata. Five of the 11 individuals showed multiple trauma. Ten (15% of MNI) individuals exhibited a total of 10 perimortal cranial traumata. This is a surprisingly high frequency of deadly head injuries, taking into account that most of the gladiator types wore helmets. A possible explanation could be the frequently reported deathblow technique used by the hammer-carrying death god "Dis Pater". The gladiator weaponry is well known through historical sources. At least one injury per known type of offensive weapon could be identified, as well as evidence for the most popular, the gladiator trident, which was found to be represented by one perimortem and two antemortem injuries. Overall the reportedly very strict nature of combat rules for gladiator fights could be confirmed by the absence of multiple perimortal traumatized individuals, showing a lack of the excessive violence commonly observed on medieval battle ground victims. This graveyard gives the opportunity to confirm historical aspects and to check the reliability of forensic methods for identification of antemortem, perimortem, or postmortem bone lesions. Typical examples for these kinds of lesions and injuries are presented. The colour of the margins of the traumata proved to be the most distinguishing feature for postmortal lesions.

Sex estimation using the femur of Austrians born in the 19th to the middle of the 20th century.

Sex estimation of human bones or bone fragments is crucial for the identification process of unknown bodies. Regional differences and secular changes lead to ever changing osteometric standards for different populations. This study provides femoral dimensions of Austrians born between 1822 and 1949 and evaluates the discriminating power of sex estimation functions developed by discriminant analyses. Depending on the state of preservation up to 127 femora of adult individuals (72 females and 55 males) were measured. The following means and standard variations were obtained: Maximum length (male: 449.7 ± 16.7 mm, female: 413.9 ± 16.3 mm), head circumference (148.5 ± 7.8 vs. 133.8 ± 6.4 mm), vertical (46.9 ± 2.4 vs. 41.9 ± 2.1 mm), transverse (46.3 ± 2.4 vs. 41.5 ± 2.0 mm), and maximum head diameter (47.0 ± 2.3 vs. 42.0 ± 2.1 mm), as well as sagittal (28.3 ± 2.2 vs. 26.2 ± 1.8 mm), transverse (27.9 ± 2.5 vs. 26.1 ± 2.0 mm), and maximum midshaft diameter (29.2 ± 2.1 vs. 27.3 ± 1.6 mm) and finally condylar width (79.6 ± 4.6 vs. 71.9 ± 3.8 mm). The (cross-validated) discriminant analysis for single measurements showed that the best classification is obtained using femur head dimensions, with correct sexing rates between 84.8 % for the circumference and 87.8 % for the maximum diameter. The maximum length achieved a similarly high rate of correct classification with 86.5 %, whereas the rate for the condylar width (80.6 %) was somewhat lower. The potential of midshaft dimensions for sex estimation was substantially weaker (62.3 to 70.8 %). With regard to a multivariate analysis, a stepwise selection procedure favoured a combination of maximum length and vertical head diameter with 90.1 % being correctly classified. A combination of maximum length, maximum head diameter and maximum midshaft diameter resulted in an even higher rate of 91.5 %. These Austrian standards may provide additional possibilities for sexing unknown bodies and skeletal remains born in the 19(th) to the middle of the 20(th) century.

A probable case of congenital syphilis from pre-Columbian Austria.

This study examines the skeletal remains of a subadult from an archeological site in Austria. Radiocarbon dating and archeological attribution indicate that this individual is of pre-Columbian origin. Most of the skeleton was recovered, and only the teeth and the orbital roofs show changes. Dental defects such as the mulberry molar and a tapered, fang-like canine suggest a diagnosis of congenital syphilis. This is the first probable case of congenital syphilis from pre-Columbian Central Europe. Our findings contribute to the pre-Columbian theory, offering counter evidence to the assumption that syphilis was carried from Columbus' crew from the New to the Old World.

Survival after trepanation-Early cranial surgery from Late Iron Age Switzerland.

Trepanation is defined as the intentional perforation of the cranial vault with removal of a piece of skull bone. In Europe, trepanation is known to have been practiced at least since the Neolithic, and it can still be found today in East African native tribes. Two skulls with lesions from the Late Iron Age site Münsingen-Rain (420-240 BC) were investigated. The aim of this study was to analyse the lesions and to determine whether they were caused by surgical interventions. Both individuals were analysed by current morphologic-anthropological methods and radiological examinations were performed with a multislice CT-scanner. Additionally, this work surveys trepanations reported in Switzerland and calculates survival rates. In Switzerland, 34 individuals with trepanations have been published. As a tendency, the survival rate appears to be relatively high from the Neolithic to Late Antiquity but then decreases until Pre-Modern times. The 78% survival rate in Late Iron Age Switzerland indicates that the surgery was often performed successfully. Skull injuries sustained in conflicts could have been a reason for trepanation during the Iron Age.

Three-dimensional histomorphometric analysis of distraction osteogenesis using an implanted device for mandibular lengthening in sheep.

The aim of this study was to lengthen the sheep mandible with a fully buried device and to quantitatively analyze the tissue regenerate in the distraction gap by means of two-dimensional and three-dimensional histomorphometry. A custom-made device for continuous distraction was used in five adult sheep and fixed with three bicortical screws on either side of an osteotomy, anterior to the premolar region of the mandible. A cable-connected power and control unit was implanted in the neck region. After a 5-day latency period, distraction was activated every 2 hours and advanced at a rate of 1.01 mm per day. The distraction period was planned for 14 days, but because of stability problems and cable breakage, the actual distraction period ranged from 2 to 17 days, resulting in gap distances from 1.7 to 17.1 mm (mean, 0.95 mm/day). Osteogenesis was followed by radiographic imaging, and after a 6-week consolidation period, the harvested mandibles were serially sectioned for histologic and two-dimensional histomorphometric analysis, with three-dimensional reconstruction. Histologic examination of the specimens demonstrated predominantly membranous bone formation with remodeling bridging the distraction gap mainly in the periosteal region of the lingual side. In addition, cartilaginous areas and chondral bone formation were observed where the bridging appeared incomplete. Because of device fixation on the buccal side of the mandible, the preservation of the lingual periosteum seemed to play the major role for sufficient bone repair in the distraction gap. Cartilage within the distraction gap suggests fixation instability in this animal model.

Citrate content of bone for time since death estimation: results from burials with different physical characteristics and known PMI.

A recently introduced method to determine the postmortem interval (PMI) based on quantification of the citrate content in bone was applied on the temporal bones and femora of 20 individuals buried in wooden coffins (WO) and body bags (BB), respectively. Concerning known vs. calculated PMI, a significant difference between the temporal and the femur bone samples of the same individuals was observed in the BB group (p = 0.012). In contrast, differences were insignificant for the WO group (p = 0.400). Moreover, similar levels of underestimation of PMIs resulted from the analysis of the femora for both burial groups (p = 0.247). Also, there was consistently less citrate preserved in the flat temporal bones as compared to the femora, indicating that the cortical layer of the long bones should be preferentially employed for citrate-based PMI estimations. The results call for additional research on subsurface-buried and surface-deposited remains to enhance the accuracy of the published PMI equation.