Age assessment in unaccompanied minors: assessing uniformity of protocols across Europe.

Age assessment of migrants is crucial, particularly for unaccompanied foreign minors, a population facing legal, social, and humanitarian challenges. Despite existing guidelines, there is no unified protocol in Europe for age assessment.The Forensic Anthropology Society of Europe (FASE) conducted a comprehensive questionnaire to understand age estimation practices in Europe. The questionnaire had sections focusing on the professional background of respondents, annual assessment numbers, requesting parties and reasons, types of examinations conducted (e.g., physical, radiological), followed protocols, age estimation methods, and questions on how age estimates are reported.The questionnaire's findings reveal extensive engagement of the forensic community in age assessment in the living, emphasizing multidisciplinary approaches. However, there seems to be an incomplete appreciation of AGFAD guidelines. Commonalities exist in examination methodologies and imaging tests. However, discrepancies emerged among respondents regarding sexual maturity assessment and reporting assessment results. Given the increasing importance of age assessment, especially for migrant child protection, the study stresses the need for a unified protocol across European countries. This can only be achieved if EU Member States wholeheartedly embrace the fundamental principles outlined in EU Directives and conduct medical age assessments aligned with recognized standards such as the AGFAD guidelines.

Analysis of interrater reliability in age assessment of minors: how does expertise influence the evaluation?

Nowadays, the clinical forensic medical management of migration flows comprises the age assessment of unaccompanied minors. The process of age estimation is a fundamental pillar for legally ensuring the minors' rights and their protection needs. The procedure is complex and involves different phases and actors, from medical doctors to law enforcement officers. The present study aimed to investigate the performance of Greulich and Pyle, Demirjian, and Mincer methods when performed by raters both trained and without training. Also, the interrater reliability within groups of raters from different areas of expertise was evaluated. A total of 36 participants were enrolled for this study, divided in two groups according to their level of experience with age estimation methods. Each rater was asked to allocate stages and standards for age assessment, evaluating ten orthopantomograms and ten hand-wrist roentgenograms. The interrater reliability expressed through the Fleiss Kappa coefficient and the agreement with the reference standard were calculated. The results showed that none of the categories analyzed could reach a good interrater reliability (ĸ > 0.8) for both methods. The study results highlighted variation and disagreement in the interpretation of the sample among raters and in the subsequent stages and standards allocation. In conclusion, the results of this study highlight that expertise does influence the reliability of the most utilized methods of age estimation of living individuals and stress the importance of proper training and practice, which could greatly increase the accuracy of age assessments.

Sex estimation of skeletons in middle and late adulthood: reliability of pelvic morphological traits and long bone metrics on an Italian skeletal collection.

There are several metric and morphological methods available for sex estimation of skeletal remains, but their reliability and applicability depend on the sexual dimorphism of the remains as well as on the availability of preserved bones. Some studies showed that age-related changes on bones can cause misclassification of sex. The purpose of this study was to establish the reliability of pelvic morphological traits and metric methods of sex estimation on relatively old individuals from a modern Italian skeletal collection. The data for this study were obtained from 164 individuals of the Milano CAL skeletal collection and average age of the samples was 75 years. In the pelvic morphological method, the recalibrated regression formula of Klales and colleagues (2012), pre-auricular sulcus, and greater sciatic notch morphology were used for sex estimation. With regard to the metric method, 15 standard measurements from upper and lower limbs were analyzed for sexual dimorphism. The results showed that in pelvic morphological approach, the application of regression formula of the revised Klales and colleague formula (2017) resulted in 100% accuracy. Classification rates of metric methods vary from 75.19 to 90.73% with the maximum epiphyseal breadth of proximal tibia representing the most discriminant parameter. This study confirms that the effect of age on sex estimation methods is not substantial, and both metric and morphological methods of sex estimation can be reliably applied to individuals of Italian descent in middle and late adulthood.

Histomorphometric analysis of osteocyte lacunae in human and pig: exploring its potential for species discrimination.

In recent years, several studies have focused on species discrimination of bone fragments by histological analysis. According to literature, the most consistent distinguishing features are Haversian canal and Haversian system areas. Nonetheless, there is a consistent overlap between human and non-human secondary osteon dimensions. One of the features that have never been analyzed for the purpose of species discrimination is the osteocyte lacuna, a small oblong cavity in which the osteocyte is locked in. The aim of this study is to verify whether there are significant quantitative differences between human and pig lacunae within secondary osteons with similar areas. Study sample comprises the midshaft of long bones (humerus, radius, ulna, femur, tibia, and fibula) of a medieval human adult and a juvenile pig. Sixty-eight secondary osteons with similar areas have been selected for each species and a total of 1224 osteocyte lacunae have been measured. For each osteon, the total number of lacunae was counted, and the following measurements were taken: minimum and maximum diameter, area, perimeter, and circularity of nine lacunae divided between inner, intermediate, and outer lacunae. Statistical analysis showed minimal differences between human and pig in the number of lacunae per osteons and in the minimum diameter (P > 0.05). On the contrary, a significant difference (P < 0.001) has been observed in the maximum diameter, perimeter, area, and circularity. Although there is the need for further research on different species and larger sample, these results highlighted the potential for the use of osteocyte lacunae as an additional parameter for species discrimination. Concerning the difference between the dimensions of osteocyte lacunae based on their position within the osteon (inner, intermediate, and outer lacunae), results showed that their size decreases from the cement line towards the Haversian canal both in human and pig.

Histomorphological analysis of the variability of the human skeleton: forensic implications.

One of the fundamental questions in forensic medicine and anthropology is whether or not a bone or bone fragment is human. Surprisingly at times for the extreme degradation of the bone (charred, old), DNA cannot be successfully performed and one must turn to other methods. Histological analysis at times can be proposed. However, the variability of a single human skeleton has never been tested. Forty-nine thin sections of long, flat, irregular and short bones were obtained from a well-preserved medieval adult human skeleton. A qualitative histomorphological analysis was performed in order to assess the presence of primary and secondary bone and the presence, absence and orientation of vascular canals. No histological sections exhibited woven or fibro-lamellar bone. Long bones showed a higher variability with an alternation within the same section of areas characterized by tightly packed secondary osteons and areas with scattered secondary osteons immersed in a lamellar matrix. Flat and irregular bones appeared to be characterized by a greater uniformity with scattered osteons in abundant interstitial lamellae. Some cases of "osteon banding" and "drifting osteons" were observed. Although Haversian bone represent the most frequent pattern, a histomorphological variability between different bones of the same individual, in different portions of the same bone, and in different parts of the same section has been observed. Therefore, the present study has highlighted the importance of extending research to whole skeletons without focusing only on single bones, in order to have a better understanding of the histological variability of both human and non-human bone.

3D quantitative analysis of early decomposition changes of the human face.

Decomposition of the human body and human face is influenced, among other things, by environmental conditions. The early decomposition changes that modify the appearance of the face may hamper the recognition and identification of the deceased. Quantitative assessment of those changes may provide important information for forensic identification. This report presents a pilot 3D quantitative approach of tracking early decomposition changes of a single cadaver in controlled environmental conditions by summarizing the change with weekly morphological descriptions. The root mean square (RMS) value was used to evaluate the changes of the face after death. The results showed a high correlation (r = 0.863) between the measured RMS and the time since death. RMS values of each scan are presented, as well as the average weekly RMS values. The quantification of decomposition changes could improve the accuracy of antemortem facial approximation and potentially could allow the direct comparisons of antemortem and postmortem 3D scans.

A comparative analysis of microscopic alterations in modern and ancient undecalcified and decalcified dry bones.

OBJECTIVES: The present study aims to evaluate the preservation of the microstructure of skeletal remains collected from four different known burial sites (archaeological and contemporary). Histological analysis on undecalcified and decalcified thin sections was performed in order to assess which of the two techniques is more affected by taphonomic insults. MATERIALS AND METHODS: A histological analysis was performed on both undecalcified and decalcified thin sections of 40 long bones and the degree of diagenetic change was evaluated using transmitted and polarized light microscopy according to the Oxford Histological Index (OHI). In order to test the optical behavior of bone tissue, thin sections were observed by polarized light microscopy and the intensity of birefringence was evaluated. RESULTS: The more ancient samples are generally characterized by a low OHI (0-1) with extensive microscopic focal destruction; recent samples exhibited a better preservation of bone micromorphology. When comparing undecalcified to decalcified thin sections, the latter showed an amelioration in the conservation of microscopic structure. As regards the birefringence, it was very low in all the undecalcified thin sections, whereas decalcification process seems to improve its visibility. DISCUSSION: The preservation of the bone microscopic structure appears to be influenced not only by age, but also by the burial context. Undecalcified bones appear to be more affected by taphonomical alterations, probably because of the thickness of the thin sections; on the contrary, decalcified thin sections proved to be able to tackle this issue allowing a better reading of the bone tissue.

Histological determination of the human origin from dry bone: a cautionary note for subadults.

Anthropologists are frequently required to confirm or exclude the human origin of skeletal remains; DNA and protein radioimmunoassays are useful in confirming the human origin of bone fragments but are not always successful. Histology may be the solution, but the young subadult structure could create misinterpretation. Histological tests were conducted on femur and skull of 31 human subjects. Each sample was observed focusing on presence or absence of fibrous bone, lamellar bone, radial lamellar bone, plexiform bone, reticular pattern, osteon banding, Haversian bone, primary osteons, secondary osteon and osteon fragments. Samples were divided into five age classes; 1 (<1 year), 2 (1-5 years), 3 (6-10 years), 4 (11-15 years) and 5 (16-20 years). Regarding femurs, class 1 presented the following: 87.5% fibrous bone, 37.5% plexiform bone, 12.5% reticular pattern and 12.5% lamellar bone radially oriented. Class 2 showed 37.5% of fibrous bone, 12.5% of reticular pattern and 37.5% of osteon banding. In the higher age classes, the classical human structures, lamellar bone and osteons were frequently visible, except for one case of reticular pattern, generally considered a distinctive non-human structure. The situation appeared different for the skull, where there was a lack of similar information, both in human and non-human. An analysis of the percentage of lamellar bone and osteons was conducted on femur and skull fragments. A trend of increase of primary osteon number and a decrease of the lamellar bone area has been detected in the femur. The present study has therefore shed some light on further pitfalls in species determination of subadult bone.

Correlation of soft tissue and skeletal injuries in cases of violent death: A retrospective study of autopsy cases for forensic anthropology.

In forensic pathology, deaths due to mechanical injuries (blunt, sharp, and gunshot) require an autopsy in order to determine the cause and manner of death through a detailed examination of viscera, soft tissues, and the skeletal system. Sometimes, such as in cases of severe putrefaction, only the skeleton is observable. In such cases, the absence of bony defects does not mean that a trauma-related death should not be considered: yet, how often will a lethal mechanical death produce a defect in the skeletal system? The present study involved a retrospective review of autopsy reports to investigate the frequencies of soft tissue and related skeletal injuries in cases of violent deaths. A total of 200 autopsy reports for cases of four mechanisms of injury including sharp force trauma (n = 50), gunshot (n = 50), blunt force trauma (n = 50), and mechanical asphyxia (n = 50) were examined. Manner of death, localization of bone fractures, and correspondence between external injuries and bone fractures were reported. Frequencies of bone fractures and degree of correspondence with soft tissue injuries for each trauma group were evaluated. The result showed that lethal blunt trauma always caused bone fractures; 94.0% of violent deaths by gunshot involved bones; 48.0% of sharp force soft tissue injuries involved the bone tissue; and hyoid bone fractures were observed in 20.0% of cases of mechanical asphyxia.

Stable Isotope Provenance of Unidentified Deceased Migrants-A Pilot Study.

In the global migration crisis, one of the challenges in the effort to identify deceased migrants is establishing their region of origin, which facilitates the search for ante mortem data to be compared with the post mortem information. This pilot study explores the potential of using stable isotope analysis to distinguish between individuals coming from West Africa and the Horn of Africa. Six individuals (four of known origin and two of unknown origin) were sampled. δ13CVPDB(keratin), δ15NVPDB(keratin) and δ18OVSMOW(keratin) of hair were analysed using Elemental Analyzers coupled with Isotope Ratio Mass Spectrometry (IRMS). δ18OVSMOW(carbonate) and δ13CVPDB(carbonate) of bone were analysed using GasBench II with IRMS, while 87Sr/86Sr composition was determined in bone and dental enamel using laser ablation multi-collector inductively coupled plasma mass spectrometry. The stable isotope compositions of the individual from the Horn of Africa differed from the other individuals. The differences found between 87Sr/86Sr of enamel and bone and between δ18O and δ13C in bone and hair reflect changes in sources of food and water in accordance with regionally typical migration journeys. The analysis of multiple stable isotopes delivered promising results, allowing us to narrow down the region of origin of deceased migrants and corroborate the information about the migration journey.

Evidence of possible lower limb amputation in a tomb in an ancient Egyptian necropolis: the case report of an on-site radiographic analysis.

Objective: Despite the existing literature, the use of surgery to treat medical diseases in Ancient Egypt remains controversial. Regarding amputations, such procedures were performed in Egypt for therapeutic reasons, although they were never described in medical papyri. Here, we present the radiographic study of a possible lower limb amputation found in a recently discovered tomb in Aswan, Egypt. Methods: The necropolis is located on the west bank of the Nile, around the Mausoleum of the Aga Khan III. Around 45 mummies were found in the tomb, along with more than 400 mixed bones of adults and subadults. Radiographic analyses were carried out directly on-site in the proximity of the tomb using a digital portable device.Among the mixed bones, we found a mature right femur with evidence of mid-diaphyseal bone interruption and exuberant reparative callus at the broken stump with woven periosteal new bone, indicating a recent and active healing process at the time of death. The X-ray study confirmed the presence of a mid-diaphyseal transverse fracture, highlighting the relatively sharp margins which were suggestive of a transverse cut at this point. A slightly radiopaque bone callus was visible as osseous spurs with circumferential and proximal directions; the exuberant bone callus revealed an ante-mortem trauma, suggesting the hypothesis of certain types of amputation. The X-ray showed no clear signs of other bone diseases or advanced taphonomic processes. Among the commingled remains, we also found the mature distal epiphysis of a right femur with radiographic evidence of extensive bone remodeling at the proximal broken stump. However, we cannot ascertain that these two femoral pieces corresponded to the same individual. Conclusions: Further studies will better clarify the causes of the bony lesion, which may be related to possible amputation of fracture from high-force blunt trauma. At present, the most likely cause relies on interpersonal violence, accidental occupation trauma and/or therapeutic treatment. Our report highlights how conventional radiology can still provide important results in the field of paleopathology, thanks to the possibility of using portable radiological devices directly on archaeological sites, thus overcoming technical difficulties in transporting bone mummified remains.

A Pilot COVID-19 Surveillance Program at the Zendrini Center in Milan (Italy) for Unaccompanied Foreign Minors.

During the COVID-19 pandemic, not only crowded refugee camps and immigration detention centers, but also receptions were places in which outbreaks occurred. To date there has been no report of the application of a COVID-19 surveillance system in reception centers for unaccompanied foreign minors only, who most of all deserve the utmost attention. Aware of this critical issue, we implemented a pilot COVID-19 surveillance program at the Zendrini center in Milan. It was started in September 2021 and was carried out for 4 months. Nasopharyngeal antigenic swabs were adopted. One day a week, two forensic physicians performed the first antigenic swab to minors who had just entered the center, or a monitoring swab after 15 days to those who were still hosted at the center. Operators were also swabbed for surveillance. A total of 80 subjects were enrolled and divided into 68 (72.5%) unaccompanied foreign minors and 22 (27.5%) operators. A total of 178 antigenic nasopharyngeal swabs were performed and tested negative. Regarding the monitoring activities, it was found that the minimum number of swabs per subject was 1 and the maximum number was 7, with an average value of 2.2 per individual. Having been able to confirm the absence of SARS-CoV-2 within the community represented a way to protect individual and collective health that could not have been pursued otherwise. Only inclusive approaches can allow communities and societies to respond more effectively to this crisis, and reduce the risk of future ones, intended as both upcoming COVID-19 waves and new infectious diseases.

Age estimation in the living: A scoping review of population data for skeletal and dental methods.

Age estimation of living individuals has become a crucial part of the forensic practice, especially due to the global increase in cross-border migration. The low rate of birth registration in many countries, hence of identification documents of migrants, especially in Africa and Asia, highlights the importance of reliable methods for age estimation of living individuals. Despite the fact that a number of skeletal and dental methods for age estimation have been developed, their main limitation is that they are based on specific reference samples and there is still no consensus among researchers on whether these methods can be applied to all populations. Though this issue remains still unsolved, population information at a glance could be useful for forensic practitioners dealing with such issues. This study aims at presenting a scoping review and mapping of the current situation concerning population data for skeletal (hand-wrist and clavicle) and dental methods (teeth eruption and third molar formation) for age estimation in the living. Two hundred studies on the rate of skeletal maturation and four hundred thirty-nine on the rate of dental maturation were found, covering the period from 1952 and 2020 for a total of ninety-eight countries. For most of the western and central African countries there are currently no data on the rate of skeletal and dental maturation. The same applies to the countries of the Middle East, as well as the eastern European countries, especially as regard the skeletal development.

Histomorphometric analysis of the variability of the human skeleton: Forensic implications.

In the last decades, the histomorphometric analysis of bone tissue has been utilized to develop equations for species discrimination of fragmentary bone. Although this technique showed promising results, its main limitation concerns the lack of knowledge on the histomorphometric variability which may exist between different bones of the skeleton. In a previous study, we demonstrated a significant histomorphological variability in different bones of the same individual and even in different sections of the same bone. The present study aimed at investigating the extent of intra-individual variability in bone histomorphometry throughout the human adult skeleton and areas of a single bone. Samples were taken along an entire medieval male adult human skeleton (aged between 26 and 45 years), including long, flat, irregular and sesamoid bones for a total of 49 cross-sections. The histomorphometric analysis revealed that the size of both Haversian systems and Haversian canals were statistically significantly larger in long and irregular bones compared to flat bones. Moreover, osteons were generally bigger in the diaphysis compared to the proximal and distal metaphyses, whereas Haversian canals showed a higher uniformity in the different portions of each bone. The present study has highlighted the importance of conducting similar studies on both human and nonhuman skeletons at different stages of skeletal maturity in order to shed light on the extent of variability in the size of osteons and Haversian canals. This, in fact, represents an important prerequisite to develop reliable histological methods for species discrimination of fragmented bone.

The Utility of Skeletal and Surgical Features for the Personal Identification Process: A Pilot Study.

This pilot study provides a conceptual framework for the application of the anthropological analysis of skeletal features and surgical interventions for the purpose of identification in cases of unknown deceased individuals with unavailable fingerprint, genetic or odontological antemortem data. The study sample includes 276 individuals with known demographic and clinical information from the Italian CAL Milano Cemetery Skeletal Collection. In the sample, 124 (45%) individuals showed one or more skeletal features that may be potentially individualizing. Of these, 79% showed two and more features, which occurred in a multitude of different combinations. Skeletal findings may provide useful postmortem information that can be compared with antemortem witness statements and clinical imaging. However, more research into the utility of dry bone findings and the availability of comparative material, including imaging, and epidemiological data needs to be undertaken before skeletal features can be implemented into identification protocols and databases.

Three-dimensional facial anatomy evaluation: Reliability of laser scanner consecutive scans procedure in comparison with stereophotogrammetry.

Laser scanner devices are acquiring a growing importance in facial anatomy. Most studies have analysed facial scans obtained through two simultaneous captures, whereas the same result can be obtained by consecutive three-dimensional (3D) scans. However, this latter procedure has not yet been validated. Fourteen volunteers underwent face image capture through stereophotogrammetry (VECTRA M3) and three consecutive facial scans through a laser scanner (Konica Minolta Vi910). The concordance between 14 linear distances, 12 angles, facial surface area and volume measurements was verified by the Bland-Altman test and calculation of absolute and relative technical errors of measurement (TEM/rTEM). The two facial images obtained by the different devices were then registered to calculate point-to-point distance. Most of linear distances and angles showed a high agreement, with "very good" or "good" rTEMs, ranging between 1.1% and 6.4%. Surface area measurements agreed well between the devices (rTEM: 6.3%), while volumes were poorly comparable (rTEM: 25.8%); the root mean square point-to-point distance was 0.80 mm (SD: 0.41). This study first tested the concordance of measurements on facial images obtained by stereophotogrammetry and consecutive laser scans. Results highlight the reliability of linear distances, angles and surface areas measurements, but discourage volume assessment and registration of surfaces acquired through different devices.

3D-3D facial superimposition between monozygotic twins: A novel morphological approach to the assessment of differences due to environmental factors.

Distinction of one twin with respect to the other, based on external appearance, is challenging; nevertheless, facial morphology may provide individualizing features that may help distinguish twin siblings. This study aims at exposing an innovative method for the facial assessment in monozygotic twins for personal identification, based on the registration and comparison of 3D models of faces. Ten couples of monozygotic twins aged between 25 and 69 years were acquired twice by a stereophotogrammetric system (VECTRA-3D® M3: Canfield Scientific, Inc., Fairfield, NJ); the 3D reconstruction of each person was then registered and superimposed onto the model belonging to the same person (self-matches), the corresponding sibling (twin-matches) and to unrelated participants from the other couples (miss-matches); RMS (root mean square) point-to-point distances were automatically calculated for all the 220 superimpositions. One-way ANOVA was used to evaluate the differences among miss-matches, twin-matches and self-matches (p < .05). RMS values for self-matches, twin-matches and miss-matches were respectively 1.0 mm (SD: 0.3 mm), 1.9 mm (0.5 mm) and 3.4 mm (0.70 mm). Statistically significant differences were found among the three groups (p < .01). Comparing RMS values in the three groups, mean facial variability in twin siblings was 55.9% of that assessed between unrelated persons and about twice higher than that observed between models belonging to the same individual. The present study proposed an innovative method for the facial assessment of twin siblings, based on 3D surface analysis, which may provide additional information concerning the relation between genes and environment.

The Issue of Age Estimation in a Modern Skeletal Population: Are Even the More Modern Current Aging Methods Satisfactory for the Elderly?

The main idea behind age assessment in adults is related to the analysis of the physiological degeneration of particular skeletal structures with age. The main issues with these procedures are due to the fact that they have not been tested on different modern populations and in different taphonomic contexts and that they tend to underestimate the age of older individuals. The purpose of this study was to test the applicability and the reliability of these methods on a contemporary population of skeletal remains of 145 elderly individuals of known sex and age. The results show that, due to taphonomic influences, some skeletal sites showed a lower survival. Therefore, the methods with the highest percentage of applicability were Lovejoy (89.6%) and Rougé-Maillart (81.3%), followed by Suchey-Brooks (59.3%), and those with the lowest percentage of applicability were Beauthier (26.2%) and Iscan (22.7%). In addition, this research has shown how for older adults the study of both acetabulum and auricular surface may be more reliable for aging. This is also in accordance with the fact that auricular surface and the acetabulum are the areas more frequently surviving taphonomic insult.

Sex Assessment from the Volume of the First Metatarsal Bone: A Comparison of Linear and Volume Measurements.

Sexual dimorphism is a crucial characteristic of skeleton. In the last years, volumetric and surface 3D acquisition systems have enabled anthropologists to assess surfaces and volumes, whose potential still needs to be verified. This article aimed at assessing volume and linear parameters of the first metatarsal bone through 3D acquisition by laser scanning. Sixty-eight skeletons underwent 3D scan through laser scanner: Seven linear measurements and volume from each bone were assessed. A cutoff value of 13,370 mm3 was found, with an accuracy of 80.8%. Linear measurements outperformed volume: metatarsal length and mediolateral width of base showed higher cross-validated accuracies (respectively, 82.1% and 79.1%, raising at 83.6% when both of them were included). Further studies are needed to verify the real advantage for sex assessment provided by volume measurements.