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.

Papaver somniferum in seventeenth century (Italy): archaeotoxicological study on brain and bone samples in patients from a hospital in Milan.

In this paper, we present the results of toxicological analyses of preserved brain tissue and bone samples from the remains of the seventeenth century patients of the Ospedale Maggiore, the main hospital in Milan and one of the most innovative hospitals in Europe from the Renaissance period. Beneath it, the crypt functioned as the burial place for the deceased of the hospital. In this multidisciplinary study of the remains, toxicological analyses in particular were performed with HPLC-MS/MS on different biological samples from nine individuals. Anthropological, paleopathological, histological, radiological examinations and radiocarbon dating were also carried out. As a result, archeotoxicological analyses revealed the presence of codeine, morphine, noscapine and papaverine, derived from Papaver somniferum, a plant present in the hospital pharmacopeia used as a narcotic, analgesic, astringent, coagulant, and antitussive agent. Such analyses have shed light on the pharmacological therapies administered to the patients near the time of death and have implemented our knowledge of medical treatment and drug administration in the 1600's.

The diachronic trend of female and male stature in Milan over 2000 years.

Stature is a biological trait directly determined by the interaction of genetic and environmental components. As such, it is often evaluated as an indicator for the reconstruction of skeletal biological profiles, past health, and social dynamics of human populations. Based on the analysis of 549 skeletons from the CAL (Collezione Antropologica LABANOF), a study of the diachronic trend of male and female adult stature in Milan (Italy) is being proposed here, covering a time span of about 2000 years, ranging from the Roman era to present-days. The skeletons, from necropolises dedicated to the less wealthy classes of Milanese society, were assigned to one of following five historical periods: Roman Era (first-fifth centuries AD), Early Middle Ages (sixth-tenth centuries AD), Late Middle Ages (eleventh-fifteenth centuries AD), Modern Era (sixteenth-eighteenth centuries AD) and Contemporary Era (nineteenth-twentieth centuries AD), and their stature was estimated according to the regression formulae of Trotter (1970). The collected data were then subjected to statistical analyses with ANOVA using R software. Although stature values showed an ample standard deviation in all periods, statistical analyses showed that stature did not significantly vary across historical periods in Milan for both sexes. This is one of the rare studies showing no diachronic changes in the trend of stature in Europe.

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.

Physical disability in Late Antiquity Milan: slipped capital femoral epiphysis with severe secondary joint disease in the Basilica of San Dionigi.

The paper presents the skeletal remains of an adult male of 30-40 years with bone lesions and deformity on the left hip, recovered during the archaeological excavation below the former Basilica of San Dionigi, dated to Late Antiquity (3rd - 5th century AD) Milan. Biological profile and paleopathological analysis were performed following standard references and the bones underwent radiological examination. Differential diagnosis included congenital anomaly, active rickets, infectious diseases, femoral neck trauma, Legg-Carvé-Perthes disease, Slipped Capital Femoral Epiphysis (SFCE), osteogenesis imperfecta and osteoporosis. While the lesions were highly consistent with Legg-Calvé-Perthes disease, they were typical of SFCE, complicated by an avascular necrosis and secondary osteoarthrosis. The alteration of the femoral head led to a 7.8% leg-length discrepancy, causing gait alteration with partial compensation though increased muscular activity on the right leg, reduced mobility of the joint and potentially hip pain, difficulty in walking and running and even limping. This paper explores a case of physical disability from the Roman era found near a Christian place of worship and represents a rare case of SFCE in the paleopathological record.

Utility of micro-CT for dating post-cranial fractures of known post-traumatic ages through 3D measurements of the trabecular inner morphology.

Fracture dating is an issue at the forefront of forensic sciences. While dating fracture is crucial to understanding and verifying the chronology of events in cases of abuse and violent death, its application is the subject of considerable discussion in the scientific community, filled with limitations and difficulties. Current methods for fracture dating are mainly based on a qualitative assessment through macroscopy, microscopy, and imaging and subject to variations depending on the experience of the observer. In this paper, we investigated the potential of quantifiable micro-CT analysis for fracture dating. Five histomorphometric parameters commonly used for the study of the 3D bone trabecular microarchitecture with micro-CT were calculated based on nine fractures of known post-traumatic ages, including the degree of anisotropy, connectivity density, bone volume fraction, trabecular thickness, and trabecular separation. As a result, trends in the evolution of the microarchitecture of the bone relative to age of the callus could be identified, in particular concerning anisotropy, trabecular separation and connectivity density, consistent with the healing bone process. The findings obtained in this pilot study encourage further research in quantifiable parameters of the bone microarchitecture as they could represent useful features for the construction of objective models for fracture dating.

How Do Drugs Affect the Skeleton? Implications for Forensic Anthropology.

Forensic anthropologists rely on a number of parameters when analyzing human skeletal remains to assist in the identification of the deceased, predominantly age-at-death, sex, stature, ancestry or population affinity, and any unique identifying features. During the examination of human remains, it is important to be aware that the skeletal features considered when applying anthropological methods may be influenced and modified by a number of factors, and particular to this article, prescription drugs (including medical and non-medical use) and other commonly used drugs. In view of this, this paper aims to review the medical, clinical and pharmacological literature to enable an assessment of those drug groups that as side effects have the potential to have an adverse effect on the skeleton, and explore whether or not they can influence the estimation of age-at-death, sex and other indicators of the biological profile. Moreover, it may be that the observation of certain alterations or inconsistencies in the skeleton may relate to the use of drugs or medication, and this in turn may help narrow down the list of missing persons to which a set of human remains could belong. The information gathered from the clinical and medical literature has been extracted with a forensic anthropological perspective and provides an awareness on how several drugs, such as opioids, cocaine, corticosteroids, non-steroidal anti-inflammatory drugs, alcohol, tobacco and others have notable effects on bone. Through different mechanisms, drugs can alter bone mineral density, causing osteopenia, osteoporosis, increase the risk of fractures, osteonecrosis, and oral changes. Not much has been written on the influence of drugs on the skeleton from the forensic anthropological practitioner perspective; and this review, in spite of its limitations and the requirement of further research, aims to investigate the current knowledge of the possible effects of both prescription and recreational drugs on bones, contributing to providing a better awareness in forensic anthropological practice and assisting in the identification process of the deceased.

Twenty years of femicide in Milan: A retrospective medicolegal analysis.

Femicide constitutes a leading cause of premature deaths for women, yet it has been the subject of limited research until recently. Enhanced data collection and analysis on killings of women and girls are necessary to understand and address this unrelenting phenomenon. This study examines all cases of female homicide encountered at the Institute of Legal Medicine of Milan (Italy) spanning from 1999 to 2019; data from 2020/2021 were shown separately given the bias that the forced cohabitation and stay at home during the lockdowns of the SARS-CoV-2 pandemic may represent regarding violence against women and femicide. In this study, specific factors were considered, including the age and nationality of the victims, the place of recovery of the bodies, the victim's relationship to the perpetrator and the injuries they suffered. As a result, 200 female killings were found among the over 15,000 autopsies and 535 homicides investigated at the Institute of Legal Medicine of Milan from 1999 to 2019, representing an average of 9.5 femicides yearly. The majority of victims were Italian (74%) and half were aged between 18 and 49 years old. The killings were overwhelmingly committed in the domestic setting (78.5%) by male perpetrators (at least 85%), related to the victims as intimate or ex-intimate partners and members of the family (73.5%). The homicides were mainly perpetrated with sharp (32%) or blunt instruments (21.5%), shooting (18.5%) and asphyxiation (16.5%). This study is part of a growing effort to enhance data collection and analysis on femicide. Studying and monitoring the rates of femicide (or "femicide watch") will permit to better understand, reduce, and finally end femicide globally.

Morphological analysis of lingula shape in a modern Italian cemeterial population: Clinical and forensic considerations.

The lingula is a small bony projection emerging from the medial ridge of the mandibular foramen, subjected to morphological variations. To date, scientific literature has described four different shapes (truncated, triangular, nodular and assimilated) and the relative distributions in different human populations. However, no data are available on Europeans so far. To this purpose, the present study aims to evaluate the distribution of the lingula shapes in the Italian population and to verify its relevance in ancestry estimation. Lingula was analysed in 235 dry mandibles selected among contemporary Italian cemeterial skeletons. Since only well-preserved sides were considered, 453 sides were evaluated according to a classification method, which includes the description of a fifth shape (the bridge shape) and the presence of mixed morphologies. In our sample, the most prevalent shape was the truncated shape (38.6%), followed by the nodular (26.3%), mixed morphologies (15.2%), triangular (10.8%), bridge (5.1%) and assimilated (4.0%) shapes. Within mixed morphologies, the most prevalent were the nodular/truncated (31.9%), nodular/triangular (30.4%), and nodular/assimilated (23.2%). Differently to previous studies, the lingula morphology could not actually offer a concrete and reliable help to ancestry estimation. However, its shape and extension would assume a stronger clinical influence for the execution and success of the alveolar nerve block used in dental and maxillofacial surgery procedures.

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.

Osteophytes: The product of convergent evolution.

The very reasonable suggestion, that diarthrodial joint and juxta-discal (vertebral centra-marginal) bony overgrowths (referred to as osteophytes) have different etiologies, has eluded previous confirmation. The prevailing perspective is that diarthrodial osteophytes represent the product of compressive forces and that those on the margins of vertebral centra result from traction and therefore are enthesial in derivation. If diarthrodial joint osteophytes result from intrinsic pressures, any surface responses would require transcortical nutritional support, easily recognized by en face microscopic examination. This contrasts with enthesially derived growth, the surface of which is characterized by Sharpey's fiber insertions. These are recognized as inverted cones with a central protrusion on examination of related bone surfaces. We hypothesize that diarthrodial and disc-adjacent osteophytes have a different pathophysiology, distinguishable on the basis of microscopic surface appearance. We pursued microscopic examination of the surfaces of osteophytes present on diarthrodial joints (hip, knee, elbow, costovertebral) and vertebrae (cervical, thoracic, and lumbar) from the CAL Milano Cemetery Skeletal Collection for presence of transcortical channels and the inverted cones of Sharpey's fiber insertions. Examination of 22 diarthrodial joint osteophytes reveals the presence solely of transcortical channels, while examination of 35 vertebral centra marginal osteophytes reveals the presence only of inverted cones. Findings are independent of age, gender, joint affected, position in the spinal column and osteophyte "severity." It is now evidenced that all osteophytes are not created equal. Diarthrodial joint osteophytes are endochondrally derived; vertebral centra osteophytes, enthesial in derivation. Different pathophysiology at least partially explain the clinical character of these processes.

The potential of micro-CT for dating post-cranial bone fractures: a macroscopic, radiographic, and microtomography study of fractures of known post-traumatic ages.

The estimation of the post-traumatic survival time (PTST) in case of bone injuries remains a tricky issue in the forensic field, especially when dealing with dry bones. Newer high-resolution imaging, and in particular microcomputed tomography (micro-CT), has the potential to significantly improve our abilities to interpret antemortem and perimortem lesions and accurately date fractures in a less destructive analysis. In this paper, nine costal fractures of known post-traumatic ages were analyzed through gross examination, conventional radiography, and microcomputed tomography, in order to test the potential of microcomputed tomography for dating fractures. As a result, microcomputed tomography provided images of high quality and definition and allowed the observation of the internal microarchitecture of the fractures and calluses. While microcomputed tomography cannot substitute histological examination for the estimation of the post-traumatic survival time, it constitutes a potent and helpful complementary tool for the analysis of bone trauma.

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.

Frequency of biological non-skeletal materials in dry bone scenarios.

In forensic anthropology, many small biological non-osseous materials may be found associated to skeletonized remains and can provide crucial information to the biological profile. Indeed, their careful recovery and correct identification can add information related to age-at-death, sex, ancestry, health, diet, migration and habits, and may be used in specific analysis such as DNA typing. However, and despite their potential, little is known about these non-osteological materials. Indeed, how frequent are they in dry bone cases? To answer this question, 100 skeletal remains with postmortem intervals ranging from 23 to 76 years from the CAL Milano Cemetery Skeletal Collection were meticulously examined for the presence of non-skeletal materials and in particular ossified cartilage, vascular calcifications, hair and nails. As a result, non-skeletal materials were found in 81 skeletal remains and showed high frequencies of recovery, despite the less-than-optimal conditions of exhumation of the skeletons. The high frequencies of non-skeletal materials obtained demonstrate the need for additional training and research given their informative power in forensic cases.

Detecting drugs in dry bone: a pilot study of skeletal remains with a post-mortem interval over 23 years.

In decomposed or skeletonized bodies, conventional matrices used in forensic toxicology may no longer be available for analysis. The aim of this paper was to test the survival and detection of toxicological substances in dry bone samples with over 23 years of post-mortem interval. In this perspective, bone samples from the cranium, ribs, and vertebrae of seven skeletons from the CAL Milano Cemetery Skeletal Collection, buried for over 23 years, fully decomposed and altered by taphonomic factors were selected based on their ante-mortem data, which included verified or suspected drug addictions or overdose. Qualitative and quantitative analyses were performed with Dionex™ ASE™ 350 Accelerated Solvent Extractor and Q-Exactive Orbitrap-mass spectrometry with a HPLC system. Positive results were obtained in six of the seven cases, and different psychoactive drugs (and in some cases their active metabolites) were detected, including analgesic (two opioids: methadone and buprenorphine) and anxiolytic drugs (benzodiazepines, in particular delorazepam, diazepam, nordiazepam, and lorazepam), a cannabinoid metabolite (THCCOOH) as well as metabolites of stimulants (benzoylecgonine and MDA). Consequently, this research shows that toxicological substances may be found in bone tissue after over 23 years of post-mortem interval.

Pitfalls of Computed Tomography 3D Reconstruction Models in Cranial Nonmetric Analysis.

Many studies in the literature have highlighted the utility of virtual 3D databanks as a substitute for real skeletal collections and the important application of radiological records in personal identification. However, none have investigated the accuracy of virtual material compared to skeletal remains in nonmetric variant analysis using 3D models. The present study investigates the accuracy of 20 computed tomography (CT) 3D reconstruction models compared to the real crania, focusing on the quality of the reproduction of the real crania and the possibility to detect 29 dental/cranial morphological variations in 3D images. An interobserver analysis was performed to evaluate trait identification, number, position, and shape. Results demonstrate a false bone loss in 3D models in some cranial regions, specifically the maxillary and occipital bones in 85% and 20% of the samples. Additional analyses revealed several difficulties in the detection of cranial nonmetric traits in 3D models, resulting in incorrect identification in circa 70% of the traits. In particular, pitfalls included the detection of erroneous position, error in presence/absence rates, in number, and in shape. The lowest percentages of correct evaluations were found in traits localized in the lateral side of the cranium and for the infraorbital suture, mastoid foramen, and crenulation. The present study highlights important pitfalls in CT scan when compared with the real crania for nonmetric analysis. This may have crucial consequences in cases where 3D databanks are used as a source of reference population data for nonmetric traits and pathologies and during bone-CT comparisons for identification purposes.

The potential of bone disease for personal identification: a case of tuberculosis.

In the forensic anthropology practice, bone diseases are rarely considered for personal identification. In this paper, we present a forensic skeletonized case with tuberculous bone lesions, for which bone pathology may provide an indicator for positive personal identification. Antemortem hospital records were available. Postmortem CT scans of the pathologically affected bones were performed, and 3D reconstructions with Global Illumination Reconstruction software (GIR) were realized, in order to confront antemortem and postmortem data. As a result, the juxtaposition and superimposition of antemortem and postmortem images evidenced several points of correspondence in the position, anatomical contour, character, and morphological characteristics of the bone lesions, thus demonstrating through a concrete case study the potential of morphological features of bone lesions for the personal identification of unknown deceased.

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.