A novel method for sorting and reassociating commingled human remains using deviation analysis

This study provides an innovative and novel method for sorting commingled human remains at the sacroiliac joint using deviation analyses. Virtual models were created at the University of Tennessee-Knoxville Donated Skeletal Collection from 69 os coxae and 66 sacra using an EinScan-Pro 2× + Handheld Surface Scanner. The shape of the auricular surfaces was analyzed in Geomagic Wrap 2017, and the congruency of the two auricular surfaces was measured using a deviation analysis. ROC curves were performed on a reference sample composed of 200 commingled and non-commingled joint pairs to identify threshold values that could help sort the commingled remains. A validation sample of 225 pairs was subsequently analyzed to demonstrate the efficacy of this new method on a sample of unknown individuals. Statistical analyses demonstrated that the deviation analysis values from sacroiliac joints of commingled pairs were significantly larger than those from non-commingled individuals (p < 0.0001). Based on the selected threshold values, 98%–100% of pairs were correctly sorted and reassociated. This novel and objective technique improves upon previously subjective strategies for sorting commingled remains and, in the future, will be applied to additional joint surfaces.     

Determining Postmortem Interval Using Glycoproteinous Adhesion Deposits by Balanus improvisus on Human Skeletal and Dental Remains

An anthropological analysis was conducted on skeletal and dental remains brought to the Galveston County Medical Examiner's office. The skeletal remains were dry, fragmented, and absent of typical fluvial characteristics. During microscopic examination, semitransparent, circular objects were discovered on the dentition, the mandible, tibial plateau, and distal femur. The objects were glycoproteinous adhesions deposited by the acorn barnacle, Balanus improvisus. B. improvisus is an intertidal barnacle found in estuaries in Galveston Bay. Basal diameter of the adhesions on the dentition were significantly smaller than those found on the postcranial bones (p = 0.010), indicating two consecutive cohorts adhered to the bone and dentition. As settlement typically occurs once a year, this would indicate that the remains were in the fluvial environment for at least 375–410 days. It is important in geographic areas that have prevalent fluvial environments that human remains, particularly dentition, are microscopically examined for marine life evidence.     

Metric Assessment of the Pubic Bone Using Known and Novel Data Points for Sex Estimation,

Biological sex estimation of skeletal remains is essential in forensic and archaeological analyses. Anthropologists most often use the pelvis, which is the most sexually dimorphic element both morphologically and metrically. While nonmetric pubic bone features have been studied extensively, few metric studies have examined this individual bone for dimorphism. For this study, three observers examined three previously identified and ten novel measurements of the pubic body on a modern sample of isolated pubic bones from the Maricopa County Forensic Science Center (FSC), in Phoenix, Arizona (n = 400). A relationship between pubic body measurements and biological sex was demonstrated, with significant correlations. Discriminant function analyses found that five measurements, four of which were novel, discriminated between males (89%) and females (86%). Observer experience level did not significantly impact the results. These five measurements were reliable and show promise for inclusion in metric methods for assessment of sex.     

A Statistical Method for Reassociating Human Tali and Calcanei from a Commingled Context

In a commingled context, assessing that a talus and a calcaneus correspond to the same individual could become a primary step for accurately sorting human remains. For this purpose, the lengths and widths of the trochlea, posterior calcaneal articular surface, and posterior talar articular surface were measured in 197 individuals (105 males, 92 females) from the Athens Collection. A total of 12 highly accurate equations for reassociating tali and calcanei were developed, using simple and multiple linear regression analysis and they were found to be suitable for sorting commingled human remains. Bilateral asymmetry and sex did not have an effect on the accuracy of the method.     

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.     

DNA Identification of Commingled Human Remains from the Cemetery Relocated by Flooding in Central Bosnia and Herzegovina

The floods in Bosnia and Herzegovina in May 2014 caused landslides all over the country. In the small village of ?eri?i, near the town of Zenica, a landslide destroyed the local cemetery, relocated graves, and commingled skeletal remains. As the use of other physical methods of identification (facial recognition, fingerprint analysis, dental analysis, etc.) was not possible, DNA analysis was applied. DNA was isolated from 20 skeletal remains (bone and tooth samples) and six reference samples (blood from living relatives) and amplified using PowerPlex^® Fusion and PowerPlex^®Y23 kits. DNA profiles were generated for all reference samples and 17 skeletal remains. A statistical analysis (calculation of paternity, maternity, and sibling indexes and matching probabilities) resulted in 10 positive identifications. In this study, 5 individuals were identified based on one reference sample. This has once again demonstrated the significance of DNA analysis in resolving the most complicated cases, such as the identification of commingled human skeletal remains.     

Bone Mineral Density Adult Age Estimation in Forensic Anthropology: A Test of the DXAGE Application

Estimating age‐at‐death of individuals represented only by skeletonized human remains is a fundamental aspect of forensic anthropological casework. Recently, several researchers have proposed that bone mineral density (BMD) is a useful predictor of age‐at‐death in forensic contexts. Navega et al. (JFS 63(2):497–503) developed an online application called DXAGE for calculating age‐at‐death from BMD parameters. This study tests the utility of DXAGE by utilizing data from the National Health and Nutrition Examination Survey (NHANES). BMD data from a female subsample (n = 470) of the NHANES 2007–2008 dataset were analyzed, and the relationship between predicted age and real age was examined. Inaccuracy was 14.25 years, and bias was ?7.20 years. Results show that there is a weak correlation between predicted and actual age (r = 0.47) using the DXAGE application. While BMD data are potentially useful for predicting age age‐at‐death, the DXAGE application should be used cautiously in forensic anthropological contexts.     

Evaluation of the Enhanced Computational Methods of Estimating Age-at-Death Using the Pubic Symphyses of a White South African Population

To address problems of reproducibility related to current age estimation methods, the enhanced computational Methods (ECM) were previously developed using a sample of white North American male pubic symphyses and showed promise in improving current techniques. However, given the evidence of sex and population differences in the onset of age markers, this study set out to test the ECM on a white South African sample. The sample consisted of 184 well-preserved os coxae from individuals of known age and sex. Pubic symphyseal surfaces were scanned using the Artec Spider 3D scanner, processed using Artec Studio 10 and analyzed using forAge. Point estimates of age were then compared to the true age of each individual. Results indicate that the ECM performed poorly in a white South African population, with consistent underestimation of age-at-death and weak positive correlations with true age. Despite the low correlations, the ECM did, however, reduce observer error.     

Advances in Osteometric Sorting: Utilizing Diaphyseal CSG Properties for Lower Limb Skeletal Pair-Matching

Pair-matching of bilateral elements is a major component of resolving commingled remains both in forensic and bioarchaeological contexts. This study presents a new method of osteometric pair-matching of the lower limbs which relies on 3D digital models of the femur and tibia bones. The proposed method, which is accompanied by a freely available open-source implementation, automatically computes a number of osteometric variables including cross-sectional geometric properties from an assemblage of left and right bone antimeres and calculates probabilistically the appropriate matching pairs as well as single elements, whose bones antimere is not present in the given assemblage. The method has been extensively tested on a skeletal sample comprising 396 femurs and 422 tibias from the Athens collection. Our results in testing commingled assemblages with no disparity show that the method’s sensitivity is 1 for sorting femurs and 0.997 for sorting tibias, whereas in assemblages with moderate disparity the sensitivity is 0.999 and 0.992 respectively. Our results further indicate that sensitivity is unaffected by the size of the commingled assemblage although the percentage of identified true matching pairs drops as the number of commingled elements increases. This means that all identified antimeres matched to an individual are still very accurately sorted despite not every individual being identified in very large assemblages. The proposed method can facilitate the sorting process of commingled remains both accurately and efficiently, while leaving a very small percentage of unsorted elements that may require further techniques for further individualization.     

Multimethod Resolution of a Small‐Scale Case of Commingling

This study highlights the use of multiple methods for resolving a case of commingled human remains. Skeletal remains were located in a marijuana field in rural northern California by law enforcement. Although initially buried in shallow graves, the remains of two decedents were disturbed by large carnivores and scattered over a wide area. The remains were submitted by law enforcement for forensic anthropological analysis and resolution of commingling. To segregate the remains of the two individuals, a number of methods were employed, including: (i) physical matching of fragmented remains; (ii) articulation to evaluate joint congruence; (iii) visual pair‐matching of bilateral elements; (iv) osteometric pair‐matching; (v) evaluation of taphonomic patterns; (vi) DNA analysis; and (vii) portable X‐ray spectrometry (pXRF). This multimethod approach resulted in accurate resolution of the commingling and facilitated reconstruction of the biological profiles, taphonomic patterns, and trauma analysis for each individual.     

Reanalysis of the Trotter Tibia Quandary and its Continued Effect on Stature Estimation of Past‐Conflict Service Members

Forensic casework from past‐conflicts relies on the corrected historical Trotter data for stature estimation in Fordisc. For roughly 10 years’, stature estimation using this data has produced point estimates for the tibia that are on average 1.25 inches less than the other long bones. This issue was identified after applying the equations derived from Fordisc to the USS Oklahoma commingled assemblage. Reevaluation of Fordisc revealed that a correction factor of 20 mm, instead of 10 mm, was mistakenly applied to the Trotter tibia data. Historical forensic anthropology reports written at the Defense POW/MIA Accounting Agency were utilized to identify that the overcorrection is isolated to Fordisc 3 with an error rate of 5% of known antemortem statures falling outside of the prediction intervals that relied on the tibia. Further evaluation of the Oklahoma sample indicates the 10 mm correction is still producing point estimates less than the other long bones.     

Potential Use of Bacterial Community Succession in Decaying Human Bone for Estimating Postmortem Interval,,

Bacteria are taphonomic agents of human decomposition, potentially useful for estimating postmortem interval (PMI) in late‐stage decomposition. Bone samples from 12 individuals and three soil samples were analyzed to assess the effects of decomposition and advancing time on bacterial communities. Results indicated that partially skeletonized remains maintained a presence of bacteria associated with the human gut, whereas bacterial composition of dry skeletal remains maintained a community profile similar to soil communities. Variation in the UniFrac distances was significantly greater between groups than within groups (p < 0.001) for the unweighted metric and not the weighted metric. The members of the bacterial communities were more similar within than between decomposition stages. The oligotrophic environment of bone relative to soft tissue and the physical protection of organic substrates may preclude bacterial blooms during the first years of skeletonization. Therefore, community membership (unweighted) may be better for estimating PMI from skeletonized remains than community structure (weighted).     

The Use of Femoral Neck Axis Length to Estimate Sex and Ancestry

Having multiple reliable methods of estimating sex and ancestry from various skeletal features increases the likelihood of identifying skeletal remains. Femoral neck axis length (FNAL), as measured in living individuals, has been shown to vary by sex and ancestry. FNAL has not, however, been previously measured directly from skeletonized remains and investigated for its potential use in forensic anthropological applications. This research proposes a method for measuring FNAL from skeletal remains, determines the reliability and repeatability of the measurement, and assesses the validity of FNAL in sex and ancestry estimation. Results showed low interobserver error in the measurement of FNAL (TEM = 0.33 mm, R = 0.99). Significant differences in FNAL were found between sexes as well as between American Black, American White, and Native American groups. FNAL can correctly classify sex in ~86% of all cases and is considered valuable to sex estimation. The value of FNAL to ancestry estimation, however, is considered limited.     

The Erratic Behavior of Lesions in Burnt Bone

This study analyses depressed fractures (by blunt force trauma) and circular full‐thickness injuries (drill injuries and gunshot wounds) in charred bones. Fifty bovine ribs (total 104 lesions) were divided into three groups. The first group consisted in 20 depressed hammer‐produced fractures; in the second one, 60 round drill‐holes were produced (30 circular, 30 semicircular); in the third group, 12 fleshed and 12 skeletonized ribs were hit by 9‐mm bullets. Each specimen was carbonized in an electric oven up to 800°C. Morphological and metric analyses were performed before and after: morphological features were preserved, but depressed fractures showed an increase in their dimensions (p‐value < 0.05); the drilled holes shrunk (p‐value < 0.01); the charring cycle increased the number of fractures in samples with gunshot wounds differently in fleshed and defleshed ribs. This study showed the complex behavior of charred bone, for what concerns the interpretation of trauma and how caution should be applied.     

The Effect of Time on Bone Fluorescence: Implications for Using Alternate Light Sources to Search for Skeletal Remains

Bones fluoresce when exposed to certain wavelengths of shortwave light, and this property can be useful in locating and sorting skeletal remains in forensic contexts. The proteins in bone collagen are largely responsible for its fluorescent properties, but these proteins degrade and denature over time. This study examined the fluorescence of bones from four temporal groups (recent, semi‐recent, ancient, and historic) ranging from 0 to 1064 years before present. Specimens were photographed under 490 nm wavelength light, and fluorescence was quantified by converting intensity to a gray scale value based on the RGB color model using ImageJ^® software. Significant (p < 0.05) differences were found in mean fluorescence between all four temporal groups, and a 0.324 coefficient of correlation indicates a significant (inverse) relationship between fluorescence and time. Bone fluorescence decreases with time, but some fluorescence is retained even in older samples. Fluorescence can therefore be reliably used in many modern skeletal remains searches.     

The Power of Exclusion using Automated Osteometric Sorting: Pair‐Matching,

This study compares the original pair‐matching osteometric sorting model (J Forensic Sci 2003 ;48:717) against two new models providing validation and performance testing across three samples. The samples include the Forensic Data Bank, USS Oklahoma, and the osteometric sorting reference used within the Defense POW/MIA Accounting Agency. A computer science solution to generating dynamic statistical models across a commingled assemblage is presented. The issue of normality is investigated showing the relative robustness against non‐normality and a data transformation to control for normality. A case study is provided showing the relative exclusion power of all three models from an active commingled case within the Defense POW/MIA Accounting Agency. In total, 14,357,220 osteometric t‐tests were conducted. The results indicate that osteometric sorting performs as expected despite reference samples deviating from normality. The two new models outperform the original, and one of those is recommended to supersede the original for future osteometric sorting work.     

Sex Determination from the Second Cervical Vertebra: A Test of Wescott’s Method on a Modern American Sample*

Abstract: Numerous methods for establishing a biological profile exist; however, many of these methods rely on the recovery of several specific bones or on fragile skeletal elements that are sometimes irrecoverable. It is for this reason new methods utilizing other previously under‐documented bones should be established and tested by the forensic anthropological community. This study tests the accuracy of Wescott’s (J Forensic Sci 2000;45(2)) method for determining sex from the second cervical vertebra. Specimens were drawn from the donated skeletal collection curated at the Hamilton County Forensic Center (n = 57) and the William M. Bass Donated Skeletal Collection (n = 243). Both intra‐ and inter‐observer error rates were low and accurate classifications ranged from 78% (females‐Function 1) to 90.6% (males‐Function 5). Of the five functions, Function 4 achieved the highest overall accuracy, with 260 individuals (86.7%) falling into the correct category. Overall, this method is an effective classificatory tool for sex estimation.     

Comparative Analysis of Insect Succession Data from Victoria (Australia) Using Summary Statistics versus Preceding Mean Ambient Temperature Models

Minimum postmortem interval (mPMI) can be estimated with preceding mean ambient temperature models that predict carrion taxon pre‐appearance interval. But accuracy has not been compared with using summary statistics (mean ± SD of taxon arrival/departure day, range, 95% CI). This study collected succession data from ten experimental and five control (infrequently sampled) pig carcasses over two summers (n = 2 experimental, n = 1 control per placement date). Linear and exponential preceding mean ambient temperature models for appearance and departure times were constructed for 17 taxa/developmental stages. There was minimal difference in linear or exponential model success, although arrival models were more often significant: 65% of linear arrival (r² = 0.09–0.79) and exponential arrival models (r² = 0.05–81.0) were significant, and 35% of linear departure (r² = 0.0–0.71) and exponential departure models (r² = 0.0–0.72) were significant. Performance of models and summary statistics for estimating mPMI was compared in two forensic cases. Only summary statistics produced accurate mPMI estimates.     

Applying Knowledge of Species‐Typical Scavenging Behavior to the Search and Recovery of Mammalian Skeletal Remains

Forensic investigations involving animal scavenging of human remains require a physical search of the scene and surrounding areas. However, there is currently no standard procedure in the U.K. for physical searches of scavenged human remains. The Winthrop and grid search methods used by police specialist searchers for scavenged remains were examined through the use of mock red fox (Vulpes vulpes) scatter scenes. Forty‐two police specialist searchers from two different regions within the U.K. were divided between those briefed and not briefed with fox‐typical scavenging information. Briefing searchers with scavenging information significantly affected the recovery of scattered bones (χ² = 11.45, df = 1, p = 0.001). Searchers briefed with scavenging information were 2.05 times more likely to recover bones. Adaptions to search methods used by searchers were evident on a regional level, such that searchers more accustom to a peri‐urban to rural region recovered a higher percentage of scattered bones (58.33%, n = 84).     

Use of 2D:4D Digit Ratios to Determine Sex

Second to fourth digit ratios (2D:4D) are sexually dimorphic in human hands and established by the 13th gestational week. Application of 2D:4D for determining sex in living individuals by Kanchan et al. (Forensic Sci Int, 181, 2008, 53.e1) produced classification rates of 80% for males and 74–78% for females. Few studies have explored the use of 2D:4D for sexing skeletal remains. We test estimated finger lengths, phalanx lengths, and 2D:4D derived from hand bones for determining sex. Maximum phalanx length was collected using a mini‐osteometric board from 451 individuals of known age, sex, and ancestry in four skeletal collections. Logistic regression of 2nd and 4th digit finger and phalanx lengths produced classification rates greater than 80%. Digit ratios, however, failed to reach classification rates greater than 59%. Our results support those of Voracek (Forensic Sci Int, 185, 2009, e29) and suggest that 2D:4D may be population‐specific and thus inappropriate for universal application as a means of determining sex.