Transcending time: the forensic anthropological case study of three unidentified transgender women in Italy in the early 1990s.

In forensic anthropology, the estimation of skeletal sex results in a dichotomous outcome: male or female. This poses challenges when dealing with transgender individuals. Italy has the highest rate per capita of transgender murders, making their identification a compelling issue. This study presents three cases of unidentified skeletal remains of transgender women (MtF) that underwent autopsies in the 1990s at the Institute of Legal Medicine of Milan. The transgender status of the individuals was determined from the autopsy records which indicated the co-existence of breast implants and male genitalia. The biological profile was created by estimating sex, age, ancestry, and stature, and the bones were investigated looking for evidence of Facial Feminization Surgery (FFS). Anthropological estimations revealed that the skeletons belong to male, adults, European individuals, between 164 and 184 cm. Fragmented gelatinous masses were found in association with two skeletons, which were interpreted as remnants of breast implants. In two cases, signs of remodeling of the cortical surface were observed on the zygomatic bones, although the bone marks observed here were not specific enough to link them to FFS. Despite some limitations, this study highlights the need for greater awareness among practitioners about this limitedly addressed issue, advocating for a more inclusive forensic anthropology that strives to improve methods and interpretation of evidence for the identification of transgender individuals.

Correlation Analysis on Anatomical Variants of Accessory Foramina in the Sphenoid Bone for Oncological Surgery.

The sphenoid bone presents several anatomical variations, including accessory foramina, such as the foramen meningo-orbitale, the foramen of Vesalius, the canaliculus innominatus and the palatovaginal canal, which may be involved in tumor invasion or surgery of surrounding structures. Therefore, clinicians and surgeons have to consider these variants when planning surgical interventions of the cranial base. The prevalence of each variant is reported in the published literature, but very little information is available on the possible correlation among different variants. Here, 300 CT scans of patients (equally divided among males and females) were retrospectively assessed to investigate the presence of the foramen meningo-orbitale, the foramen of Vesalius, the canaliculus innominatus and the palatovaginal canal. Possible differences in the prevalence of each accessory foramen according to sex were assessed, as well as possible correlations among different variants through the Chi-square test (p < 0.01). Overall, the prevalence of the foramen meningo-orbitale, the foramen of Vesalius, the canaliculus innominatus and the palatovaginal canal was 30.7%, 67.7%, 14.0% and 35.3%, respectively, without any difference according to sex (p > 0.01). A significant positive correlation was found between the foramen of Vesalius and canaliculus innominatus, both in males and in females (p < 0.01). In detail, subjects with canaliculus innominatus in 85.7-100.0% of cases also showed the foramen of Vesalius, independently from sex and side. The present study provided novel data about the prevalence of four accessory foramina of the sphenoid bone in an Italian population, and a correlation between the foramen of Vesalius and the canaliculus innominatus was found for the first time. As these accessory foramina host neurovascular structures, the results of this study are thus useful for appropriate planning surgical procedures that are tailored to the anatomical configuration of the patient and for improving techniques to avoid accidental injuries in cranial base surgery. Knowledge of the topography, frequencies and the presence/absence of these additional foramina are pivotal for a successful procedure. Clinicians and surgeons may benefit from these novel data for appropriate recognition of the variants, decision-making, pre-operative and treatment planning, improvement of the procedures, screening of patients and prevention of misdiagnosis.

Rumor has it: A narrative review on the use of skeletal non-metric traits and variants for personal identification.

The human skeleton displays an immense array of traits and variant features that are elements of inter-individual variability. The general assumption is that they may represent individualizing markers for the personal identification of unidentified decedents, but very few works consider them as such. This review provides an overview on the possible use of non-metric traits and skeletal variants for personal identification. The paper discusses the issues related to unquantified comparisons, then it presents a statistical approach based on frequencies of these features for identifying unknown remains. Narrowing down an initial number of 1000 papers, the core of the review is represented by 10 papers that considered non-metric traits and skeletal variants as individualizing features, according to both qualitative and quantitative assessments. Despite visual examination remains the gold-standard, more sound methods are requested to quantify the strength of a match or a mismatch. This especially applies in the wake of juridical demands, hence also satisfying the desire of prosecutors and judges to rely on a "quantified" risk. To this purpose, non-metric traits and skeletal variants seem to be a suitable tool to provide quantified evidence, when related frequencies are known.

Assessment of Anatomical Uniqueness of Maxillary Sinuses through 3D-3D Superimposition: An Additional Help to Personal Identification.

Paranasal sinuses represent one of the most individualizing structures of the human body and some of them have been already analyzed for possible applications to personal identification, such as the frontal and sphenoid sinuses. This study explores the application of 3D-3D superimposition to maxillary sinuses in personal identification. One hundred head CT-scans of adult subjects (equally divided among males and females) were extracted from a hospital database. Maxillary sinuses were segmented twice from each subject through ITK-SNAP software and the correspondent 3D models were automatically superimposed to obtain 100 matches (when they belonged to the same person) and 100 mismatches (when they were extracted from different individuals), both from the right and left side. Average RMS (root mean square) point-to-point distance was then calculated for all the superimpositions; differences according to sex, side, and group (matches and mismatches) were assessed through three-way ANOVA test (p < 0.017). On average, RMS values were lower in matches (0.26 ± 0.19 mm in males, 0.24 ± 0.18 mm in females) than in mismatches (2.44 ± 0.87 mm in males, 2.20 ± 0.73 mm in females) with a significant difference (p < 0.001). No significant differences were found according to sex or side (p > 0.017). The study verified the potential of maxillary sinuses as reliable anatomical structures for personal identification in the forensic context.

Comparing Genetic and Physical Anthropological Analyses for the Biological Profile of Unidentified and Identified Bodies in Milan.

When studying unknown human remains, the estimation of skeletal sex and ancestry is paramount to create the victim's biological profile and attempt identification. In this paper, a multidisciplinary approach to infer the sex and biogeographical ancestry of different skeletons, using physical methods and routine forensic markers, is explored. Forensic investigators, thus, encounter two main issues: (1) the use of markers such as STRs that are not the best choice in terms of inferring biogeographical ancestry but are routine forensic markers to identify a person, and (2) the concordance of the physical and molecular results. In addition, a comparison of physical/molecular and then antemortem data (of a subset of individuals that are identified during our research) was evaluated. Antemortem data was particularly beneficial to evaluate the accuracy rates of the biological profiles produced by anthropologists and classification rates obtained by molecular experts using autosomal genetic profiles and multivariate statistical approaches. Our results highlight that physical and molecular analyses are in perfect agreement for sex estimation, but some discrepancies in ancestry estimation were observed in 5 out of 24 cases.

How Physical and Molecular Anthropology Interplay in the Creation of Biological Profiles of Unidentified Migrants.

The skeletal sex and ancestry of unidentified human crania can be inferred both from physical and from molecular features. This paper depicts and discusses the experiences of physical and molecular anthropologists on a set of commingled crania from the largest Mediterranean shipwreck disaster on 18 April 2015, in order to facilitate identification of human crania. Twenty-one disarticulated crania that were recovered from the above-mentioned shipwreck were analyzed to estimate skeletal sex and ancestry, following a physical and a molecular pipeline. The physical analyses applied morphological and metric methods that provided posterior probabilities for the crania to be classified into a sex or ancestral group. The molecular analyses were performed on petrous bones via a shotgun sequencing approach that allowed us to determine the sex of each individual and to retrieve the complete mitochondrial genome, Y chromosome single nucleotide polymorphisms, up to 597573 SNPs across the human genome from each individual. The morphometric sex analyses showed that most crania belonged to male individuals, although some estimations remained uncertain or undetermined. Inconsistent results were obtained for ancestry estimation as well, since morphological methods classified the crania mostly as European/White, in contrast to the most numerous African forms determined by craniometric analyses. This quite agreed with molecular analyses that identified only African males. Overall, undetermined and contrasting results were obtained between disciplines, preventing the creation of reliable and sound biological profiles that could provide guidance on the sex and ancestral group of the victims. Therefore, the times may not be mature for a merger of physical and molecular anthropology. However, future investigations of this research avenue would pave the way to the possible development of novel tools, methods, and wider reference databases that could address the limitations of both disciplines.

Skeletal Markers of Physiological Stress as Indicators of Structural Violence: A Comparative Study between the Deceased Migrants of the Mediterranean Sea and the CAL Milano Cemetery Skeletal Collection.

Structural violence is an indirect form of violence that can lead to physiological consequences. Interestingly, these physiological disruptions may affect the skeletons and can therefore provide relevant information on violence and way of life in the analysis of skeletal remains. The aim of the present study was to test the hypothesis that migrants who died in the Mediterranean Sea would present physiological cranial stress markers such as cribra orbitalia (CO), porotic hyperostosis (PH), and linear enamel hypoplasia (LEH) more frequently and more severely than Italians of the 20th century. With this intent, a total of 164 crania were examined: 139 from deceased migrants recovered from a shipwreck in the Mediterranean Sea in 2015, aged between 16 and 35 years old, and 25 of the same age from the CAL Milano Cemetery Skeletal Collection. Both presence and severity of CO, PH, and LEH were evaluated. The data obtained were analyzed using Wilcoxon signed-rank and independence Chi-squared tests to compare the results between the two samples and to test whether there was an association between the sample of migrants and the occurrence of lesions. As a result, CO and PH appeared more frequently and more severely in the migrant sample. In addition, migrants were significantly associated with CO, PH, and LEH (p-values < 0.05). Although this does not imply in any way that CO, PH, and LEH are specific to migration, they should be regarded as indicators of structural violence.

Digital pair-matching of iliac bones: pilot study on a three-dimensional approach with models acquired through stereophotogrammetry.

Three-dimensional (3D) pair-matching has brought about an innovative approach for the analysis of commingled skeletal remains, and it has been tested on bone models acquired through CT and laser scans. Here, 3D models of 40 innominate bones (20 left and 20 right) of 20 documented male individuals from a cemeterial skeletal collection were acquired through a stereophotogrammetric device (VECTRA M3, Canfield Scientific, Inc.). The ventral iliac surface was chosen as the anatomical region of interest (ROI) for the analysis. Each left ROI was mirrored and superimposed on the matching right ROI (contralateral element from the same individual) and mismatching ROIs (contralateral elements from different individuals). The point-to-point distance between models was calculated through the Vectra Analysis Module (VAM) software and the root mean square (RMS) point-to-point distance value was used to evaluate the sorting performance of the method, in terms of sensitivity and specificity rates. Differences in RMS between matches and mismatches were investigated through a Student's t test (p < 0.05). The state of preservation of the remains was assessed following an index of anatomical completeness and differences in RMS distances of true matches according to different anatomical completeness were assessed through the Mann-Whitney test (p < 0.05). RMS point-to-point distances of matches and mismatches were significantly different (p < 0.01), being the matches lower than mismatches. The RMS threshold of 2.9 mm identified all the true pairs; the test was 100% sensitive and 51% specific. The RMS of matches with a better state of preservation are significantly lower than the less preserved matches (p < 0.05). In general, a low RMS distance value may indicate a true match, being it to be further verified. The 3D approach for sorting innominate bones provides a valid screening test that could complete subjective and osteometric methods with numerical evidence of the match. Preliminary data suggest a possible relation between RMS distance values and taphonomic condition, which would benefit from further research.

3D-3D facial registration method applied to personal identification: Does it work with limited portions of faces? An experiment in ideal conditions.

Personal identification of faces represents a challenging issue, especially for what concerns the quantification of the comparison. The 3D-3D superimposition approach proved to distinguish between matches and mismatches. However, the potential of this procedure applied to cases where only parts of faces are visible still has to be verified. This study aimed at verifying the applicability of a 3D-3D procedure to faces divided into three thirds. 3D models of fifty male subjects acquired through stereophotogrammetry were used. The 3D facial models were divided into upper, middle, and lower thirds and registered onto other models belonging to the same and different individuals according to the least point-to-point distance. In total, 50 matches and 50 mismatches were analyzed. RMS value (root mean square) of point-to-point distance between the two facial surfaces was calculated through VAM® software. Statistically significant differences between matches and mismatches in each facial third were assessed through Mann-Whitney test (p < 0.05). On average, RMS value in matches was 0.32 ± 0.12 mm in upper third, 0.36 ± 0.15 mm in middle third, and 0.40 ± 0.20 mm in lower third, respectively; in mismatches, RMS value was 1.40 ± 0.32 mm in upper third, 1.96 ± 0.58 mm in middle third, and 2.39 ± 0.90 mm in lower third, respectively. Differences in RMS values between matches and mismatches were significantly different for all facial thirds, without superimpositions (p < 0.01). This study shows that the existing 3D-3D superimposition methods may be useful also when only a limited portion of face is visible in ideal conditions. Their application to forensic cases of identification still needs to be verified.

Taphonomic study on drowned victims in a non-sequestered aquatic environment in the Mediterranean Sea.

Human decomposition in sea water poses several challenges to forensic practitioners tasked with the analysis of drowned bodies. Postmortem changes in the marine environment have not been extensively investigated and the current literature is mainly based on bodies retrieved from shallow waters or on limited samples. On 18 April 2015, a fishing boat carrying allegedly 1,000 migrants sank in the Mediterranean Sea. In a fifteen-month time span, humanitarian missions were carried out to recover the bodies from the sea. The present study investigates postmortem changes on the drowned victims in a non-sequestered environment in the Mediterranean Sea. A retrospective study was performed by two observers on the autopsy photographic records in the series of bodies recovered from the open sea. For 184 bodies, the postmortem changes were evaluated according to facial, body, limb, and total aquatic decomposition score (FADS, BADS, LADS, TADS, respectively). Furthermore, a modification to the current scoring system that divides upper and lower limbs was suggested. The interobserver agreement was assessed using Krippendorff's alpha coefficient. Possible relations between the decomposition scores and PMSI were investigated with Pearson correlation analysis. According to the sequence of the recovery missions, increasing FADS, BADS, LADS, and TADS were observed. The modified scoring system showed a strong agreement between observers, allowing a more accurate description of the actual extent of decomposition. The scores showed a significant relation with the PMSI (p < 0.01). Prolonged submersion in the open marine environment was confirmed to show increasing decomposition rates, from moderate decay to full disarticulation. This study provides a descriptive unicum of the postmortem changes in the open sea, which may contribute to strengthen the discipline and aid description of bodies recovered in similar circumstances, especially if a body needs to be associated to a disaster or period with respect to another, thus facilitating families or authorities in the search for specific victims.

3D-3D Superimposition of Pubic Bones: Expanding the Anthropological Toolkit for the Pair-Matching of Commingled Skeletal Remains.

Virtual anthropology (VA) has recently produced an additional tool for the analysis of commingled remains and is based on the distance analysis between three-dimensional (3D) models of bones. To date, the pair-matching of the innominate bone through a 3D approach remains partially unexplored. Here, 44 abdominal CT scans (22 males and 22 females) were selected from a hospital database, and the pubic bones were segmented through ITK-SNAP software. The models were hollowed with Viewbox4 to minimize the amount of trabecular bone. The left pubic bones were mirrored and superimposed on the right ones, according to the smallest point-to-point difference between the two surfaces through VAM software. RMS distances between models were calculated through VAM, producing RMS values for 20 matches and 420 mismatches for each sex group. Differences in RMS distance values between matches and mismatches were investigated through Mann−Whitney tests (p < 0.05); the repeatability of the procedure was assessed through absolute and relative technical error measurement (TEM and rTEM). RMS distance values of matches and mismatches were significantly different (p < 0.01) in both groups. The method yielded optimal results with high sensitivity (100.0%) and specificity (99.8% in males, 98.8% in females) rates according to the chosen threshold. This project contributes to the research field of VA with a valuable adjunct that may bolster and strengthen the results of the current visual and osteometric methods through a multidisciplinary approach.

Improving 3D-3D facial registration methods: potential role of three-dimensional models in personal identification of the living.

Personal identification of the living from video surveillance systems usually involves 2D images. However, the potentiality of three-dimensional facial models in gaining personal identification through 3D-3D comparison still needs to be verified. This study aims at testing the reliability of a protocol for 3D-3D registration of facial models, potentially useful for personal identification. Fifty male subjects aged between 18 and 45 years were randomly chosen from a database of 3D facial models acquired through stereophotogrammetry. For each subject, two acquisitions were available; the 3D models of faces were then registered onto other models belonging to the same and different individuals according to the least point-to-point distance on the entire facial surface, for a total of 50 matches and 50 mismatches. RMS value (root mean square) of point-to-point distance between the two models was then calculated through the VAM® software. Intra- and inter-observer errors were assessed through calculation of relative technical error of measurement (rTEM). Possible statistically significant differences between matches and mismatches were assessed through Mann-Whitney test (p < 0.05). Both for intra- and inter-observer repeatability rTEM was between 2.2 and 5.2%. Average RMS point-to-point distance was 0.50 ± 0.28 mm in matches, 2.62 ± 0.56 mm in mismatches (p < 0.01). An RMS threshold of 1.50 mm could distinguish matches and mismatches in 100% of cases. This study provides an improvement to existing 3D-3D superimposition methods and confirms the great advantages which may derive to personal identification of the living from 3D facial analysis.

Exploring the potential of cranial non-metric traits as a tool for personal identification: the never-ending dilemma.

Forensic anthropologists tasked with identification of skeletal remains often have to set up new strategies to overcome the limitations of conventional individualizing markers. A sound acquaintance with non-metric traits is essential for a reliable distinction between normal variations and pathological or traumatic conditions, yet the role of cranial variants in the identification process is still somehow ill-defined. One hundred crania (50 males and 50 females) of known sex and age were selected from the Collezione Antropologica LABANOF (a documented contemporary skeletal collection) and non-metric traits were scored as present or absent and by side. The frequencies of 13 traits were used to calculate the compound probabilities to find an individual with an exact combination of cranial features in the worldwide population. The probabilities of the majority of the individuals (53%) are within the 1 out of 10 million-1 out of 1 million interval. However, a fair number of subjects (25%) of the sample have the probabilities falling into the 1 out of 1 billion-1 out of 100 million interval, while the probabilities of a small portion of the sample (10%) are less than 1 out of 1 billion. This pilot study illustrates that some combinations of cranial variants are quite rare and may represent potential evidence to discern presumptive identifications, when an appropriate set of traits is selected and antemortem data are available for comparison. However, further research on larger and various samples is needed to confirm or discard the use of combinations of cranial non-metric traits as individualizing markers.

Does the choice of the reference model affect the results of 3D-3D superimposition procedure? A comparison of different protocols for personal identification.

In literature, 3D-3D superimposition has been widely recognized as a valid method for personal identification. However, very little information is available about possible variability due to differences in protocols of registration of 3D models and calculation of RMS (root mean square) point-to-point distance. Frontal sinuses from 50 CT scans were segmented twice through the ITK-SNAP software and grouped in two samples (1 and 2). Maximum breadth, height and volume were measured. 3D models belonging to the same subject were then superimposed one on each other in 50 matches. In addition, superimposition of 50 random mismatches was performed. For each superimposition, the procedure was repeated four times choosing different reference models both for registration and calculation of RMS. Differences in RMS value among protocols of registration and RMS calculation were assessed through paired Student's t-test (p < 0.05). Possible correlations between differences in RMS among groups and differences in frontal sinus size between the superimposed models were analysed through calculation of Pearson's correlation coefficient (p < 0.05). Results showed that RMS calculation did not yield significant differences according to which 3D model is used as reference; on the other hand, RMS values from registration procedure significantly differ according to which model is chosen as reference, but only in the mismatch group (p < 0.001). Differences in RMS value according to RMS calculation are dependent upon all the three measurements, whereas differences according to registration protocols were significantly related only with the breadth of frontal sinuses but only in mismatches (p < 0.001). In no case, superimpositions of RMS values were found between matches and mismatches. This article for the first time proves that the protocol of registration and calculation of RMS significantly influences the results of 3D-3D superimposition only in case of mismatches.

A probable case of holoprosencephaly with cyclopia in a full-term fetus from a modern skeletal collection.

OBJECTIVE: This study presents evidence of a probable case of holoprosencephaly with cyclopia, which has been rarely reported in the paleopathological literature. MATERIALS: The skeletal remains of a male fetus between 36 and 40 gestational weeks from the Collezione Antropologica LABANOF (CAL) Milano Cemetery Skeletal Collection were studied. METHODS: The bones were macroscopically examined, and pathological anomalies were recorded and evaluated alongside paleopathological and clinical literature. RESULTS: Developmental anomalies were observed. In particular, a single orbit and optical canal were present, and the frontal, sphenoid and palatine bones were prematurely fused. These changes altered the normal morphology of the midline structures of the cranium and face. CONCLUSIONS: The developmental anomalies observed are consistent with a case of holoprosencephaly associated with cyclopia. SIGNIFICANCE: Holoprosencephaly is a fatal congenital condition caused by the failure of the prosencephalon to separate in two halves. This condition is clinically well-known, with an estimated modern incidence of 1/16,000 births; however, the paleopathological literature lacks reports that would help anthropologists and paleopathologists interpret these anomalous signs on dry bone. This report documents a rare paleopathological case of the condition on a full-term fetus from a modern skeletal collection. LIMITATIONS: Taphonomic and anthropic factors may have impaired the observation of all pathological features. SUGGESTIONS FOR FURTHER RESEARCH: Comparative studies with cases from documented collections could improve knowledge of the appearance of this condition on dry bones.

Advances in the identification of deciduous molar tooth germs.

In forensic anthropology, correct identification of human deciduous teeth is of paramount importance for age-at-death estimation and relies on detailed anatomical descriptions. Yet literature is scarce on indications: details on the morphology of molar tooth germs of fetuses and newborns, developing from multiple mineralized centers that will eventually coalesce, are scant. This paper presents new anatomical elements for practitioners to identify human molar tooth germs at early developmental stages. 126 deciduous molars from 22 modern skeletons of fetuses and newborns (with a known age-at-death ranging between 0 days and 2 months and 21 days postnatal), without reported or observed dental pathological signs, were selected from the Collezione Antropologica LABANOF (CAL) documented skeletal collection. Gross anatomical descriptions of the morphology and configuration of the centers were provided, considering the number of mineralized centers, the shape and the outline of the occlusal plane at different stages. Three different developmental stages were observed in the maxillary first and second molar and the mandibular first molar, whereas in the mandibular second molar four stages were observed. For each stage, we provide additional detailed morphological descriptions, sketches outlining the shape of the tooth germ, and a picture of the tooth; also, indications for siding the teeth are presented. This information can be used by forensic anthropologists and odontologists for a proper identification when tooth germs are not found in anatomical connection within the dental sockets. Further analyses that encompass more age groups on a larger sample would allow to map the entire crown development of deciduous molars.