DXAGE 2.0 - adult age at death estimation using bone loss in the proximal femur and the second metacarpal.

The accurate age at death assessment of unidentified adult skeletal individuals is a critical research task in forensic anthropology, being a key feature for the determination of biological profiles of individual skeletal remains. We have previously shown that the age-related decrease of bone mineral density (BMD) in the proximal femur could be used to assess age at death in women (Navega et al., J Forensic Sci 63:497-503, 2018). The present study aims to generate models for age estimation in both sexes through bone densitometry of the femur and radiogrammetry of the second metacarpal. The training sample comprised 224 adults (120 females, 104 males) from the "Coimbra Identified Skeletal Collection," and different models were generated through least squares regression and general regression neural networks (GRNN). The models were operationalized in a user-friendly online interface at https://osteomics.com/DXAGE2/ . The mean absolute difference between the known and estimated age at death ranges from 9.39 to 13.18 years among women and from 10.33 to 15.76 among men with the least squares regression models. For the GRNN models, the mean absolute difference between documented and projected age ranges from 8.44 to 12.58 years in women and from 10.56 to 16.18 years in men. DXAGE 2.0 enables age estimation in incomplete and/or fragmentary skeletal remains, using alternative skeletal regions, with reliable results.

Estimation of the post-mortem interval in human bones by infrared spectroscopy.

In forensic anthropology, there is an inherent difficulty in estimating the post-mortem interval (PMI). This study aimed to assess whether there is a correlation between changes in the bone mineral component and the PMI estimation. Samples of femur and humerus from 80 identified individuals with known post-mortem interval were analyzed. Infrared spectroscopy in attenuated total reflectance mode (FTIR-ATR) was applied for this analysis, several indices having been obtained from the infrared bands most representative of the bone's relative contents in carbonate and phosphate. Specific relationships between sex/age and PMI were attained: for larger PMIs, there was an increase in the amount of B-type carbonate (BPI), A-type carbonates (API), and in the carbonate/phosphate (C/P) ratio, and a decrease of the crystallinity index (CI) and in the carbonate ratio (C/C). Two particular infrared indices (CI and C/C) were identified as the most suitable for post-mortem interval estimation, especially in females, controlling the effects of sex and age (in the statistical analysis).

Elemental Composition in Female Dry Femora Using Portable X-Ray Fluorescence (pXRF): Association with Age and Osteoporosis.

Pathophysiological conditions can modify the skeletal chemical concentration. This study analyzes the elemental composition in two anatomical regions from dry femoral bone using a portable X-Ray Fluorescence (pXRF) and evaluates its impact in the bone mineral density (BMD). The left femora of 97 female skeletons (21-95 years old individuals) from the Coimbra Identified Skeletal Collection were studied. Diagenetic biases were discarded at the outset and BMD was determined with Dual-energy X-ray absorptiometry. Chemical measurements were performed at the midpoint of the femoral neck and at the midshaft using a pXRF device, and comparisons were made considering the age and the BMD values. Only elements with a Technical Measurement Error ≤ 5% were selected: P, S, Ca, Fe, Zn, As, Sr, Pb and the Ca/P ratio. Statistically significant differences were found between regions, with higher concentrations of P, Ca, Zn and S at the midshaft, and the Ca/P ratio at the femoral neck. The concentration of P is higher in individuals < 50 years, while S and Ca/P ratio increase in individuals ≥ 50 years. The decrease of P with age can be simultaneously related to the decline of its concentration in osteoporosis. Decreased BMD is also associated with higher levels of S and Pb. Osteoporosis enhances the absorption of osteolytic elements in specific locations. This fast and non-destructive technique has proved effective for the comprehension of chemical changes related to bone mass loss. This study highlights the potential of identified skeletal collections to improve the knowledge about bone fragility.

New data about the 21st Century Identified Skeletal Collection (University of Coimbra, Portugal).

After the publication of the first article in 2014, 21st Century Identified Skeletal Collection, housed in the Laboratory of Forensic Anthropology, Department of Life Sciences at the University of Coimbra, Portugal, has been growing. Currently, the collection is composed of 302 complete adult skeletons of both sexes, which means that in 5 years it has doubled. The collection consists mostly of elderly individuals, with only 12.25% of the individuals aged less than 61 years old. All individuals are Portuguese nationals who died between 1982 and 2012. Ninety individuals exhibit prostheses, other medical devices and signs of surgical procedures. Moreover, a sub-collection of experimentally burned skeletons is under development, and currently includes 56 individuals (18.54% of the collection). The 21st Century Identified Skeletal Collection constitutes a fundamental tool for forensic anthropology research, including the development and validation studies of methods that focus on elderly individuals, as can be ascertained by the numerous scientific publications and academic scholarship that have been produced in previous years.

The association of osteochemometrics and bone mineral density in humans.

OBJECTIVES: Even though much is known about bone mineral and matrix composition, studies about their relationship with several bone properties and its alterations related to bone diseases such as osteoporosis are practically non-existent in humans. Thus, the development of methods to understand the effects of bone properties at a microscopic level is paramount. This research aimed to evaluate whether Fourier transform infrared-attenuated total reflectance (FTIR-ATR) band intensity ratios correlate with femoral bone mass, bone mineral content (BMC) (total and femoral neck), bone mineral per unit area (BMD) (total, femoral neck, greater trochanter, intertrochanteric region, and Ward's area) and the area (total and femoral neck). A sample of femora from the 21st Century Identified Skeleton Collection (N = 78, 42 females and 36 males) was employed and BMC, BMD, and the femoral areas were acquired by DXA. RESULTS: It was found that only females' BMD had a significant association with the femoral FTIR-ATR indices under study, whereas bone collagen (Am/P) and the content of carbonate Type A (API) in males correlated with the total proximal femur area of the regions of interest and the femoral neck area. DISCUSSION: Men and women showed different changes related to their chemical composition in BMD, BMC, and probed area, most likely due to differences in structure and physiology, as well as mechanical strength in the proximal femoral sites where BMD was analyzed.

Beyond metrics and morphology: the potential of FTIR-ATR and chemometrics to estimate age-at-death in human bone.

In forensic anthropology, the application of traditional methods for estimating the biological profile of human skeletal remains is often hampered by poor preservation and skeletal representativeness, compromising their reliability. Thus, the development of alternative methods to the morphometric analysis of bones to estimate the biological profile of human remains is paramount. The age of an individual can cause changes in bone morphology, mass and size, as well as in its chemical composition. In this sense, the main objective of this research was to evaluate if the contents of bone collagen (Am/P), carbonate type A (API), carbonate type B (BPI), the relation between the carbonate content (types A and B) to type B carbonate (C/C), carbonate-phosphate ratio (C/P) and crystallinity index (CI), spectroscopic indices obtained from relationships between infrared absorption band intensities (FTIR-ATR), can be used as age-at-death predictors. A sample of femora and humeri from the 21st Century Identified Skeleton Collection (N = 80, 44 females and 36 males) was employed. Results show that, with advancing age, women's femora have lower CI values, but BPI and C/P indices increase, and the deformation and disorder of the crystal lattice are probably affected by the integration of type B carbonate content of the femur. The ratios analysed, especially the CI and the BPI, show potential to estimate age-at-death in human skeletal remains, when sex is already known, thus helping to assess the biological profile when conventional methods cannot be applied.

A method for estimating gestational age of fetal remains based on long bone lengths.

The estimation of gestational age (GA) in fetal human remains is important in forensic settings, particularly to assess fetal viability, in addition to often being the only biological profile parameter that can be assessed with some accuracy for non-adults. The length of long bone diaphysis is one of the most frequently used methods for fetal age estimation. The main objective of this study was to present a simple and objective method for estimating GA based on the measurements of the diaphysis of the femur, tibia, fibula, humerus, ulna, and radius. Conventional least squares regression equations (classical and inverse calibration approaches) and quick reference tables were generated. A supplementary objective was to compare the performance of the new formulae against previously published models. The sample comprised 257 fetuses (136 females and 121 males) with known GA (between 12 and 40 weeks) and was selected based on clinical and pathological information. All measurements were performed on radiographic images acquired in anonymous clinical autopsy records from spontaneous and therapeutic abortions in two Portuguese hospitals. The proposed technique is straightforward and reproducible. The models for the GA estimation are exceedingly accurate and unbiased. Comparisons between inverse and classical calibration show that both perform exceptionally well, with high accuracy and low bias. Also, the newly developed equations generally outperform earlier methods of GA estimation in forensic contexts. Quick reference tables for each long bone are now available. The obtained models for the estimation of gestational age are of great applicability in forensic contexts.

One for all and all for one: Linear regression from the mass of individual bones to assess human skeletal mass completeness.

OBJECTIVES: Complete and accurate human skeletal inventory is seldom possible in archaeological and forensic cases involving severe fragmentation. In such cases, skeletal mass comparisons with published references may be used as an alternative to assess skeletal completeness but they are too general for a case-by-case routine analysis. The objective is to solve this issue by creating linear regression equations to estimate the total mass of a skeleton based on the mass of individual bones. MATERIALS AND METHODS: Total adult skeletal mass and individual mass of the clavicle, humerus, femur, patella, carpal, metacarpal, tarsal, and metatarsal bones were recorded in a sample of 60 skeletons from the 21st century identified skeletal collection (University of Coimbra). The sample included 32 females and 28 males with ages ranging from 31 to 96 years (mean = 76.4; sd = 14.8). Skeletal mass linear regression equations were calculated based on this sample. RESULTS: The mass of individual bones was successfully used to predict the approximate total mass of the adult skeleton. The femur, humerus, and second metacarpal were the best predictors of total skeletal mass with root mean squared errors ranging from 292.9 to 346.1 g. DISCUSSION: Linear regression was relatively successful at estimating adult skeletal mass. The non-normal distribution of the sample in terms of mass may have reduced the predictive power of the equations. These results have clear impact for bioanthropology, especially forensic anthropology, since this method may provide better estimates of the completeness of the skeleton or the minimum number of individuals. Am J Phys Anthropol 160:427-432, 2016. © 2016 Wiley Periodicals, Inc.

Assigned sex estimation with the clavicle and scapula: A study in a Portuguese reference sample.

The estimation of biological sex is a critical step in the assessment of the biological profile of an anonymous skeletonized individual. In certain recovery circumstances, the most dimorphic skeletal areas, such as the pelvis, are absent or fragmented; in that case, other bones of the skeleton, including the clavicle and scapula, can be used to predict sex. The purpose of this research is to generate new models for the estimation of sex with clavicular and scapular measurements using a study-sample of 129 individuals with clavicle (65 males and 64 females) and 112 individuals with scapula (50 males and 62 females) from the Lisbon Identified Skeletal Collection (Portugal). A decision tree classifier (C4.5) and logistic regression (LR) were employed to create univariable and multivariable sex prediction models. Accuracy under cross-validation of the classification models is high (up to 93.8%), with minimal bias (<5%), particularly in the multivariable models. The proposed LR models facilitate the probabilistic estimation of biological sex, accounting for the significant overlap in the expression of sexual dimorphism.

The Identification Potential of Atherosclerotic Calcifications in the Context of Forensic Anthropology.

Atherosclerosis is an inflammatory disease that, in its more developed stages, can lead to the calcification of fatty plaques on the walls of arteries, resulting in the appearance of new bone elements. It is a condition that has been studied and documented little in the context of paleopathology, especially in the framework of forensic anthropology. This article analyzed the skeletal remains of 71 individuals (35 females and 36 males) from the Luís Lopes Identified Collection of the National Museum of Natural History and Science in Lisbon, 31 of whom had an autopsy report. An attempt was made to ascertain whether these bone elements resulting from atherosclerotic calcification would resist cadaveric decomposition and whether they would be recoverable several years after burial, and a survey was carried out of their distribution according to sex and age, as well as their association with other pathologies, such as osteoporosis and cardiac and renal pathologies. An imaging analysis of an atherosclerotic plaque was also carried out to complement the macroscopic analysis and present other methods of identifying plaques. It was concluded that each atherosclerotic calcification has a unique profile, which can be useful for identification, especially in cases where the individual shows a severe condition. In terms of identification potential, the analysis of calcified atherosclerotic plaques can be useful, as they can corroborate or reject an identification. However, it always requires the existence of ante-mortem imaging exams and must always be used in addition to other identification methods. Regardless of the identification, these plaques are bone elements resulting from a pathology and should, therefore, be known and recognized by the scientific community.

A case of early childhood caries from a Medieval site in Southern Portugal: a multidisciplinary approach.

Dental caries is the most prevalent chronic infectious disease during childhood both in historical and contemporary times, but research focused on the oral health of non-adults from the past is still scant. As such, this study proposes a multidisciplinary approach to the differential diagnosis of severe dental lesions in a medieval non-adult skeleton. The skeleton of a three-year-old child recovered in the medieval necropolis of Cacela Velha (Portugal) was studied through macroscopic, radiological, elemental and stable isotope analyses. This individual exhibited enamel destruction and dentine exposure in both the maxillary and mandibular teeth, with the latter also showing changes in coloration. Elemental analysis showed that his skull presented lower values of Si, Cl, and Ca and higher of Cu compared to the control, while the concentration of P and S were significantly lower in the teeth. Early childhood caries is the most probable diagnosis for the dental lesions observed, apparently stemming from a reticulate of factors that include potential malnutrition, and the consumption of sugars in complementary feeding - even though historical sources point to the scarcity of sugar in Portugal during most of the Middle Ages.

Secular Trends in the Size and Shape of the Scapula among the Portuguese between the 19th and the 21st Centuries.

Potential secular changes in the human scapula are fundamentally unbeknownst, with most of the preceding anatomical studies focusing on long-term changes in the long bones and the skull. As such, the cardinal purpose of this study pertains to the evaluation of secular trends on the shape and size of the scapula in a time period spanning from the 19th to the early 21st centuries. The study sample included 211 individuals (100 males and 111 females) from the Coimbra Identified Skeletal Collection and the 21st Century Identified Skeletal Collection. The size and shape of the scapula were evaluated using geometric morphometrics. Results show secular changes over a relatively short period of time in both the shape and size of the scapula in Portuguese nationals. Shape changes were observed in both sexes but expressed minimally, while a significant negative trend in the size of the scapula was detected in males. Scapular size decrement in males conceivably echoes general trends of the overall anatomy towards a narrower body associated with higher standards of living that include enhanced nutrition and universal healthcare, among other factors.

Performance of the supervised learning algorithms in sex estimation of the proximal femur: A comparative study in contemporary Egyptian and Turkish samples.

Sex estimation standards are population specific however, we argue that machine learning techniques (ML) may enhance the biological sex determination on trans-population application. Linear discriminant analysis (LDA) versus nine ML including quadratic discriminant analysis (QDA), support vector machine (SVM), Decision Tree (DT), Gaussian process (GPC), Naïve Bayesian (NBC), K-Nearest Neighbor (KNN), Random Forest (RFM) and Adaptive boosting (Adaboost) were compared. The experiments involve two contemporary populations: Turkish (n = 300) and Egyptian populations (n = 100) for training and validation, respectively. Base models were calibrated using isotonic and sigmoid calibration schemes. Results were analyzed at posterior probabilities (pp) thresholds >0.95 and >0.80. At pp = 0.5, ML algorithms yielded comparable accuracies in the training (90% to 97%) and test sets (81% to 88%) which are not modified after employing the calibration techniques. At pp >0.95, the raw RFM, LDA, QDA, and SVM models have shown the best performance however, calibration techniques improved the performance of various classifier especially NBC and Adaboost. By contrast, the performance of GPC, KNN, QDA models worsened by calibration. RFM has shown the best performance among all models at both thresholds whereas LDA benefited the best from using both calibration methods at pp >0.80. Complex ML models are not necessarily achieving better performance metrics. LDA and QDA remain the fastest and simplest classifiers. We demonstrated the capability of enhancing sex estimation using ML on an independent population sample however, differences in the underlying probability distribution generated by models were detected which warranted more cautious application by forensic practitioners.

Sex assessment with the radius in Portuguese skeletal populations (late 19th - early to mid 20th centuries).

The assessment of sex is of immense relevance in the analysis of human skeletal remains, as other parameters of the biological profile are usually sex-specific (e.g., age at death or stature). The radius can be used to estimate sex when more dimorphic bones are not available or in the case of incomplete and fragmentary remains. Ten radius measurements collected in a sample of 364 individuals (166 females and 198 males) from the Coimbra Identified Skeletal Collection (late 19th - early 20th centuries) were employed to generate univariable and multivariable models for sex estimation. All models were evaluated with a 10-fold cross-validation method and an independent holdout sample from the Luís Lopes Collection (late 19th - mid 20th centuries) encompassing 50 individuals (25 females and 25 males). Univariable models show an accuracy ranging from 77.7% to 89.8% (cross-validation), and from 70% to 86% (test sample), while accuracy in the multivariable models varies from 88.7% to 93.4% (cross-validation), and 84.0% to 90.0% (test sample). Results suggest that measurements of the radius are useful to develop standard guidelines for sex estimation of anonymous skeletal remains.

Why are tapes better than wires in knotless rotator cuff repairs? An evaluation of force, pressure and contact area in a tendon bone unit mechanical model.

PURPOSE: Knotless repairs have demonstrated encouraging performance regarding retear rate reduction, but literature aiming at identifying the specific variables responsible for these results is scarce and conflictive. The purpose of this paper was to evaluate the effect of the material (tape or wire suture) and medial tendon passage (single or double passage) on the contact force, pressure and area at the tendon bone interface in order to identify the key factors responsible for this repairs´ success. METHODS: A specific knotless transosseous equivalent cuff repair was simulated using 2 tape or suture wire loaded medial anchors and 2 lateral anchors, with controlled lateral suture limb tension. The repair was performed in a previously validated sawbones® mechanical model. Testing analyzed force, pressure and area in a predetermined and constant size "repair box" using a Tekscan® sensor, as well as peak force and pressure, force applied by specific sutures and force variation along the repair box. RESULTS: Tapes generate lower contact force and pressure and double medial passage at the medial tendon is associated with higher contact area. Suture wires generate higher peak force and pressure on the repair and higher mean force in their tendon path and at the medial bearing row. Force values decrease from medial to lateral and from posterior to anterior independently of the material or medial passage. CONCLUSION: Contrary to most biomechanical literature, suture tape use lowers the pressure and force applied at the tendon bone junction, while higher number of suture passage points medially increases the area of contact. These findings may explain the superior clinical results obtained with the use uf suture tapes because its smaller compressive effect over the tendon may create a better perfusion environment healing while maintaining adequate biomechanical stability.

CADOES: An interactive machine-learning approach for sex estimation with the pelvis.

The pelvis is consistently regarded as the most sexually dimorphic region of the human skeleton, and methods for sex estimation with the pelvic bones are usually very accurate. In this investigation, population-specific osteometric models for the assessment of sex with the pelvis were designed using a dataset provided by J.A. Serra (1938) that included 256 individuals (131 females and 125 males) from the Coimbra Identified Skeletal Collection and 38 metric variables. The models for sex estimation were operationalized through an online application and decision support system, CADOES. Different classification algorithms generated high accuracy models, ranging from 85% to 92%, with only three variables; and from 85.33% to 97.33%, with all 38 variables. CADOES conveys a probabilistic prediction of skeletal sex, as well as a suite of attributes with educational applicability in the fields of human skeletal anatomy and statistics. This study upholds the value of the pelvis for the estimation of skeletal sex and provides models for that can be applied with high accuracy and low bias.

Maternal mortality, marital status and bone mineral density in young women from the Coimbra identified skeletal collection.

ABSTRACT: In contemporary populations, pregnancy and lactation are usually followed by transient bone loss. The observation of reduced bone mass in young females from archaeological sites has sometimes been interpreted as an outcome of reproductive stress. In order to evaluate the overall effect of reproductive dynamics on bone mass in a historical skeletal sample, bone mineral density (BMD) at the proximal femur was assessed by dual-energy X-ray absorptiometry in 78 young women (17-39 years) from the Coimbra Identified Skeletal Collection. BMD was compared within the skeletal sample ("maternal deaths" [ICD - 10: chapter XV] vs. "other causes of death", and "married/widowed women" vs. "single women"). Results revealed that mean BMD differences among groups are non-significant, suggesting that a strict reproductive interpretation of premature bone loss in young women from archaeological contexts is not sustained by empirical evidence. Bone mass in young women from archaeological sites should be interpreted as a complex trait stemming from the interplay between reproductive factors, genetics, nutrition, physical activity, and age at menarche.

Extracapsular fracture of the femur in an elderly male from the Church of Nossa Senhora da Anunciada (Setúbal, Portugal).

In modern populations, hip fractures in older people are associated with high morbidity and mortality. Their incidence is rising; notwithstanding, fractures of the proximal femur are still relatively uncommon in archeological contexts. This case study represents a well-healed hip fracture in an aged male skeleton from Church of Nossa Senhora da Anunciada (16th-19th centuries AD) in Setúbal (Portugal). The individual was also diagnosed with diffuse idiopathic skeletal hyperostosis. Fractures of the proximal femur are usually associated with bone loss but in this case other causes are proposed, including the anatomy of the proximal femur, and the potential combined effect of diffuse idiopathic skeletal hyperostosis, cardiovascular disease, diabetes mellitus and falls.

DXAGE: A New Method for Age at Death Estimation Based on Femoral Bone Mineral Density and Artificial Neural Networks.

Age at death estimation in adult skeletons is hampered, among others, by the unremarkable correlation of bone estimators with chronological age, implementation of inappropriate statistical techniques, observer error, and skeletal incompleteness or destruction. Therefore, it is beneficial to consider alternative methods to assess age at death in adult skeletons. The decrease in bone mineral density with age was explored to generate a method to assess age at death in human remains. A connectionist computational approach, artificial neural networks, was employed to model femur densitometry data gathered in 100 female individuals from the Coimbra Identified Skeletal Collection. Bone mineral density declines consistently with age and the method performs appropriately, with mean absolute differences between known and predicted age ranging from 9.19 to 13.49 years. The proposed method-DXAGE-was implemented online to streamline age estimation. This preliminary study highlights the value of densitometry to assess age at death in human remains.