Forensic Sciences doi: 10.3390/forensicsci6020039

Authors: Elizabeth R. Schifris Wilfredo Henriquez-Madrid Darrell D. Horton Daniel F. Gallego

Background/Objectives: Left ventricular free wall rupture is a rare but catastrophic complication of acute myocardial infarction with extremely high mortality. Deaths occurring in water environments present unique forensic challenges requiring systematic evaluation of drowning, intoxication, trauma, and natural disease. This case report describes a fatal left ventricular free wall rupture occurring shortly after successful percutaneous coronary intervention (PCI), emphasizing the medicolegal differential diagnosis and the importance of comprehensive postmortem evaluation. Results: A 58-year-old man with non-ST-elevation myocardial infarction underwent successful PCI with three drug-eluting stents and was discharged home. Six hours later, he developed severe back pain and was found unresponsive in a bathtub. Autopsy demonstrated a 2.6 cm transmural rupture of the anterolateral left ventricular free wall with 150 mL of hemopericardium. Postmortem computed tomography (PMCT), performed as part of routine forensic evaluation, had identified hemopericardium prior to autopsy. Histology showed coagulative necrosis with neutrophilic infiltration. The rupture site was remote from stented vessels with no procedural injury. Toxicology revealed therapeutic medication levels. Pulmonary and scene findings did not support drowning as a cause of death. Conclusions: Ventricular free wall rupture remains a relevant cause of sudden death following myocardial infarction despite successful revascularization. Comprehensive forensic evaluation integrating scene investigation, macroscopic autopsy findings, histopathology, and toxicology is essential to distinguish natural disease progression from accidental or iatrogenic causes in deaths occurring in water environments. This case highlights that ventricular free wall rupture can occur shortly after apparently successful PCI and underscores the importance of comprehensive forensic evaluation in water-associated deaths.

Forensic Sciences doi: 10.3390/forensicsci6020038

Authors: Muhammad Jefri Mohd Yusof Tharshini Chandran Muhammad Reza Amin Reza Adnan Eddy Saputra Rohmatul Amin Sarah Aliah Amir Sarifudin Nurul Ain Abu Bakar

Background/Objectives: Bloodstain pattern analysis (BPA) is widely used in crime scene investigation (CSI), yet its practical application, evidential limits, and interpretive role are often discussed in fragmented or technique-focused terms. This systematic literature review examines how BPA is used in CSI, with emphasis on its operational functions, interpretive scope, and scientific robustness. Methods: The review followed PRISMA 2020 guidelines. A comprehensive search was conducted in Scopus using predefined Boolean strings. After screening, eligibility assessment, and manual review, 18 peer-reviewed research articles published between 1996 and 2026 were included. Data were extracted systematically and analysed using thematic synthesis. Results: The findings show that BPA is applied in CSI as an integrated evidential pathway rather than as a single analytical procedure. Its uses include bloodstain detection and documentation, geometric reconstruction through trajectory and area-of-origin analysis, differentiation of mechanisms and sources to prevent misclassification, activity-level inference based on transfer and contact phenomena, and temporal reasoning related to trace formation. The review also highlights the role of validation infrastructures, including blood substitutes, animal analogues, and computational methods, which support training, experimentation, and reproducibility under ethical and practical constraints. Across the literature, reconstruction accuracy is shown to be sensitive to documentation quality, measurement assumptions, environmental conditions, and contextual limitations. Conclusions: Overall, BPA contributes to CSI by enabling structured, context-aware interpretation of blood evidence while remaining subject to measurement assumptions, contextual influences, and cognitive factors that may affect reconstruction outcomes. Its evidential value lies not only in reconstructing events, but also in supporting transparent, testable, and defensible forensic reasoning.

Forensic Sciences doi: 10.3390/forensicsci6020037

Authors: Paolo Bailo Maria Garreffa Giuseppe Gennari Giussy Barbara Edgardo Somigliana Lorenzo Franceschetti Andrea Piccinini

Background/Objectives: Forensic DNA analysis is a key investigative resource in sexual-violence casework, but its recorded use depends on how cases move through clinical, custody, judicial, and laboratory pathways. In Italy, this transition is organizationally heterogeneous and poorly documented through linkable procedural data. This study aimed to describe archive-based trends in archived cases, documented judicial seizures, and recorded forensic DNA analyses within a Milan-based clinical-forensic custody pathway from 2016 to 2024. Methods: A retrospective, archive-based observational study was conducted using records from the institutional UNIMI forensic custody observatory linked to the SVSeD pathway. The primary dataset included annual counts of archived sexual-violence-related cases, cases with documented judicial seizure of biological evidence, and cases with recorded forensic DNA analysis. For contextual interpretation, aggregate centre-level indicators, namely total sexual-violence cases and ex officio reports, were also reviewed as non-linkable background data. Only descriptive analyses were performed, and no causal inference was attempted. Results: Over the study period, the archive recorded 2311 cases, 102 cases with documented judicial seizure of biological evidence, and 142 cases with recorded forensic DNA analysis. The temporal series showed marked variability, including a decline in 2020, followed by recovery in subsequent years. Most recorded forensic DNA analyses were documented as being performed by the Polizia Scientifica and Carabinieri (RIS/RaCIS), whereas smaller proportions involved the university laboratory and other external laboratories. Contextual indicators showed a progressive increase in total sexual-violence cases over time, while ex officio reports did not rise proportionally. Conclusions: The findings document pathway-level attrition between archived cases, documented seizure, and recorded forensic DNA analysis, but they do not identify the exact procedural stage at which attrition occurs. The absence of standardized case-level procedural variables precludes causal attribution for non-seizure or non-analysis of biological evidence. Future evaluation of forensic DNA utilization in sexual-violence casework will require standardized tracking across clinical, custody, judicial, and laboratory stages.

Forensic Sciences doi: 10.3390/forensicsci6020036

Authors: Arianna DeCorte Gabrielle Wolfe M. Katherine Philpott Christopher J. Ehrhardt

Background/Objectives: The goal of this study was to characterize changes in autofluorescence of epithelial cells obtained from saliva stains that occur with time and investigate the potential for these changes to serve as time-since-deposition (TSD) signatures for this sample type. Methods: Saliva from 50 individuals was used to create 208 deposits that were aged between one day and nine months. Autofluorescence profiles of individual cells were obtained from each sample using imaging flow cytometry (IFC) and analyzed across nine different emission channels ranging between 435 nm and 800 nm. Results: Results showed strong evidence for linear increases in autofluorescence intensity when epithelial cells from a single donor deposit were measured over time (12 of 14 donors r ≥ 0.9). When autofluorescence profiles from all 50 donors were combined into a single time series, variation in autofluorescence intensity was observed between individual deposits with the same TSD. This inter-contributor variation decreased the overall strength of the linear relationship (r = 0.83) and yielded residual errors of ~8 days for samples that were actually 1 day old and ~82 days for samples that were over 180 days old using a linear regression model. Although this approach may not currently be amenable to estimating TSD to the day with high accuracy, clear, non-overlapping differences in autofluorescence intensity were still observed between certain time intervals, e.g., saliva deposits that were aged for 1 day compared to saliva deposits that were aged for more than 120 days. Conclusions: This suggests that cellular autofluorescence signatures have the potential to be probative when hypotheses for sample deposition involve disparate time intervals or as a screening tool for identifying which samples are most likely relevant to the crime in question based on their deposition time.

Forensic Sciences doi: 10.3390/forensicsci6020035

Authors: Suthat Duangchit Woranan Kirisattayakul Prin Twinprai Naraporn Maikong Nattaphon Twinprai Jiratcha Witchathrontrakul Thongjit Mahajanthavong Chalermphon Pitirith Kanokwan Lamai Phatthiraporn Aorachon Sararat Innoi Nareelak Tangsrisakda Sitthichai Iamsaard Chanasorn Poodendaen

Background/Objectives: Sex estimation from craniofacial morphology is a fundamental component of biological profile construction in forensic anthropology. Population-specific reference data for Thai individuals derived from computed tomography (CT) remain limited, and direct comparisons between discriminant function analysis (DFA) and machine learning classifiers are frequently complicated by inconsistent validation protocols. This study aimed to characterize sexual dimorphism in CT-derived craniofacial measurements, compare the classification performance of DFA, support vector machine (SVM), and random forest (RF) under a unified validation protocol, and demonstrate their practical application in a forensic context. Methods: CT images from 300 Thai adults (150 males, 150 females; age range 20–90 years) were obtained from Srinagarind Hospital, Khon Kaen University. Eight linear craniofacial measurements spanning the cranial vault, facial skeleton, nasal aperture, and orbital region were obtained from each case. DFA, SVM, and RF were developed and compared under a unified leave-one-out cross-validation protocol. Classification performance was assessed using accuracy, AUC, and Matthews correlation coefficient (MCC). Results: Seven of eight measurements exhibited statistically significant sexual dimorphism, with facial breadth and nasal height demonstrating the greatest dimorphism. DFA achieved the highest classification accuracy of 85.7%, AUC of 0.924, and MCC of 0.713, incorporating five measurements into the canonical function. SVM and RF achieved comparable accuracy of 84.7% and 84.0%, respectively. All three classifiers correctly classified both forensic application cases with high confidence. Conclusions: CT-derived craniofacial measurements provide a reliable basis for sex estimation in Thai adults. The convergence of performance across all three classifiers under a unified internal validation protocol strengthens confidence in the internally validated performance estimates. The derived discriminant function equation and saved machine learning models constitute a complementary and immediately applicable toolkit for CT-based forensic sex estimation in the Thai population.

Forensic Sciences doi: 10.3390/forensicsci6020034

Authors: Michal Soták Mária Chovancová Petra Švábová Zuzana Kozáková Radoslav Beňuš

Background: Latent fingerprints are crucial forensic evidence, but their stability can be affected by environmental factors such as direct sunlight. The findings indicate that prolonged sunlight exposure may be associated with reduced fingerprint quality and forensic usability. Methods: A total of 322 groomed latent fingerprints from one volunteer were deposited on non-porous glass and exposed to direct sunlight for 1–7 weeks. A control sample was preserved without exposure. Fingerprints were developed using magnetic powder and assessed by minutiae counts. Usability was classified according to Slovak forensic standards. Statistical analysis was conducted using the Friedman test and Durbin–Conover test. Results: Significant differences in minutiae counts were observed between the control and selected exposure intervals (weeks 1, 3, 4, 6 and 7; p < 0.05). The degradation pattern was not linear, with initial decreases followed by stabilization in later weeks. Despite statistical differences, 99.38% of fingerprints remained usable for identification, and none were classified as non-usable. Conclusions: Prolonged direct sunlight exposure did not substantially reduce the identificatory value of groomed latent fingerprints on glass. Even after several weeks, most fingerprints retained sufficient ridge detail for personal identification, supporting their evidential relevance in outdoor forensic contexts.

Forensic Sciences doi: 10.3390/forensicsci6020033

Authors: Chaniswara Hengcharoen Churdsak Jaikang Giatgong Konguthaithip Paknaphat Watwaraphat Karune Verochana Tawachai Monum

Background: Reliable identification remains a cornerstone of forensic investigations, particularly when encountering degraded remains or suboptimal biological evidence. This study evaluates the potential of dentine metabolomics, utilizing proton nuclear magnetic resonance (1H-NMR) spectroscopy, to detect cancer-associated metabolic signatures in dental tissues for forensic applications. Methods: Forty-four non-carious second molars were analyzed, comprising 22 samples from deceased individuals with a documented history of cancer and 22 age- and sex-matched controls. Metabolomic profiling was conducted using 1H-NMR spectroscopy to identify and quantify dentine metabolites. Statistical evaluation included unsupervised principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), receiver operating characteristic (ROC) curve analysis, and exploratory binary logistic regression. Results: Among the 209 identified metabolites, inosinic acid and 2-ketobutyric acid were identified as the most robust discriminative biomarkers across both multivariate and univariate frameworks. The exploration within-sample predictive model achieved a Nagelkerke R2 of 0.822 and an overall classification accuracy of 90.9%, with a specificity of 95.5% and a sensitivity of 86.4%. These key metabolites are fundamentally associated with purine metabolism and oxidative stress pathways frequently dysregulated in oncogenesis. Conclusions: This pilot study suggests that dentine may retain metabolomic information associated with cancer comorbidity under heterogeneous postmortem conditions. However, the findings remain exploratory and require validation in larger cohorts with standardized postmortem variables before practical forensic implementation.

Forensic Sciences doi: 10.3390/forensicsci6020032

Authors: Inês Lopes Cardoso Mariana Sá Maria Teresa Moreira Maria Inês Guimarães

Background/Objectives: The presence of postmortem pink teeth is a phenomenon occasionally observed during forensic autopsies. This phenomenon has been typically associated with deaths involving drowning, asphyxiation, trauma, or electrocution. However, the recent literature suggests that it is nonspecific. Despite being recognized in forensic literature for decades, its pathophysiology, prevalence, and forensic relevance remain controversial. The existing evidence is scattered and primarily based on case reports, narrative reviews, or isolated observations. This scoping review aims to systematically map the available scientific evidence on the phenomenon of postmortem pink teeth and its potential role in determining the cause of death within the context of forensic medicine. Methods: This review was conducted following the Joanna Briggs Institute (JBI) methodology for scoping reviews and reported according to the PRISMA-ScR (Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews) guidelines. Results: A comprehensive search was carried out across multiple databases to identify studies addressing the occurrence, mechanisms, and forensic interpretation of pink teeth in postmortem examinations. After application of the inclusion and exclusion criteria, the search allowed for the selection of 12 publications (case reports and observational studies). Conclusions: Collected data from the selected studies allowed us to conclude that PTP is a nonspecific taphonomic artifact, lacking diagnostic value in determining the cause of death. Mapping the current evidence on pink teeth contributed to a better understanding of this phenomenon and its lack of forensic significance, promoting more informed decision-making in postmortem assessments.

Forensic Sciences doi: 10.3390/forensicsci6010031

Authors: Jillian Morgan Candice Bridge

Background/Objectives: The opioid crisis is an ongoing problem in the United States, and fentanyl analogs play a major role in the issue, as novel fentanyl analogs are constantly being developed. Substitutions and additions to the fentanyl scaffold impact the potency of the substances and can sometimes influence the biotransformation of the drugs. This study aimed to determine whether one or more common metabolites could be detected among a group of five methoxylated fentanyl analogs, for the purpose of eventually providing a more rapid detection method for new and unknown fentanyl-related compounds in toxicological samples. Methods: In vitro metabolism via pooled human liver microsomes (HLMs) was performed for five methoxylated fentanyl analogs (para-methoxyfentanyl, ortho- and para-methoxy butyryl fentanyl, and ortho- and para-methoxy furanyl fentanyl) to generate metabolites. Assays were analyzed via liquid chromatography–tandem mass spectrometry. Results: Nine metabolites were detected. A common metabolite was observed between fentanyl analogs that were methoxylated at the para-position. Conclusions: Similarities between metabolites of five methoxylated fentanyl analogs were noted. It appeared that the major pathway of metabolism for methoxylated fentanyl analogs is largely consistent, regardless of the location of the methoxy substituent so long as the piperidine nitrogen is unobstructed and is available for N-dealkylation. This research provides valuable insight to potentially find new ways for toxicologists to identify novel methoxylated fentanyl analogs in biological matrices.

Forensic Sciences doi: 10.3390/forensicsci6010029

Authors: Francesco Orsini Luigi Cipolloni Paola Frati Giovanni Pollice Chiara Fabrello Stefania De Simone

Background/Objectives: Abortion legislation represents a complex intersection of medical practice, ethical considerations, and legal frameworks that demonstrate significant legal heterogeneity across Europe. This study undertakes a comprehensive comparative assessment of the statutory schemes governing abortion across the European continent, examining gestational limits, exceptional circumstances, and regulatory requirements. Methods: A comparative legal analysis was conducted across 31 European jurisdictions. Primary legislative instruments were identified and authenticated through official governmental sources, parliamentary databases, and legal repositories to ensure analysis of current consolidated legislation. Data extraction focused on gestational limits, exceptional circumstances, procedural requirements, and constitutional provisions to categorize jurisdictions into regulatory models. Additionally, a structured literature search was performed in PubMed and Scopus (2015–2025) using the keywords “abortion,” “law,” and “Europe.” From 297 screened records, 30 articles were selected to contextualize legislative evolution and scholarly discourse. Results: The comparative analysis identified substantial heterogeneity in European abortion legislation, revealing four distinct regulatory models. Most jurisdictions establish a legal limit for elective abortion of approximately 12 weeks of gestation, with variations ranging from 10 weeks to 24 weeks. Exceptions to gestational limits are widely recognized for maternal life-threatening conditions, severe fetal anomalies and pregnancies resulting from sexual violence. Conclusions: European abortion legislation reflects persistent regulatory pluralism rather than convergence toward a unified model. While commonality exists regarding early gestational limits for elective abortion, significant variation remains in exceptional circumstances, procedural requirements, and underlying regulatory philosophies. This heterogeneity impacts healthcare provision, cross-border reproductive care, and medico-legal practice. The identified regulatory models illustrate diverse balances between reproductive autonomy and state interests. Future research should examine the practical consequences of these diverse schemes on health outcomes and cross-border patient mobility.

Forensic Sciences doi: 10.3390/forensicsci6010030

Authors: Phatthiraporn Aorachon Tarinee Sawatpanich Suthat Duangchit Chanasorn Poodendaen Sitthichai Iamsaard

Background/Objectives: Calcaneal spurs are pathological bone formations at entheseal attachment sites with clinical implications but limited forensic anthropological applications. While entheseal changes have been proposed as age estimation markers in forensic contexts, empirical validation remains insufficient, particularly for Southeast Asian populations. This study evaluated calcaneal spur utility for forensic age estimation in Thai skeletal remains while establishing population-specific osteological reference data for forensic individuation. Materials and Methods: The 3516 dry calcanei from 1758 Northeastern Thai skeletons (1031 males, 727 females; age 22–106 years) were examined. Spurs were classified by anatomical location as dorsal (D-type), plantar (P-type), or combined plantar–dorsal (P–D type). The morphometric measurements were performed bilaterally. Age-associated patterns were analyzed across four age cohorts (≤40, 41–50, 51–60, ≥61 years), and Random Forest machine learning classification tested forensic age estimation capacity using 10-fold cross-validation. Results: Overall prevalence reached 67.63% with distinctive P–D type predominance. While age-stratified prevalence increased from 24.56% (≤40 years) to 74.77% (≥61 years), Random Forest modeling explicitly demonstrated overall classification accuracy of 62.5%. Compared between sexes, the maximum length of calcaneal spurs was significantly longer in males. Dimensional analyses revealed weak age correlations and substantial inter-individual morphological variation precluded reliable age prediction. Interestingly, the unique P–D type distribution pattern (77.5% among spur-bearing individuals) may serve as an auxiliary marker for Thai population affinity assessment in forensic contexts. Conclusions: This study established the first comprehensive Thai-specific osteological reference for calcaneal spurs, revealing distinctive plantar–dorsal type predominance valuable for forensic population affinity assessment and provided population-specific baseline data for forensic individuation.

Forensic Sciences doi: 10.3390/forensicsci6010028

Authors: Larisa Adela Udriştioiu Mihai Andrei

Dental hard tissues, through their remarkable resistance to degradation, represent one of the most durable biological materials available for postmortem investigation. The preparation of undecalcified or ground sections allows microscopic visualization of enamel, dentin and cementum structures, which can preserve chronological, physiological, or environmental information. This review provides a comprehensive overview of the forensic applications of dental hard tissue ground sections, focusing on methodological principles, interpretive potential and practical constraints. The literature in forensic odontology highlights their relevance for age estimation through tooth cementum annulation, identification of neonatal and accentuated stress lines, and the assessment of thermal or chemical alterations. While these methods have proven scientific validity in anthropology and histology, their forensic implementation remains limited by heterogeneity in protocols and interpretative subjectivity. Standardization of preparation techniques, digital imaging, and integration with complementary analyses such as micro-CT or SEM could enhance the reliability and medico-legal relevance of this classical but underused approach.

Forensic Sciences doi: 10.3390/forensicsci6010027

Authors: Lina De Paola Damiano Marchesini Monica La Greca Flavia Sciuttini Anna Claudia Caruso Gabriele Napoletano Bruno Cirillo Enrico Marinelli

Background/Objectives: Viper envenomation in Italy is uncommon but carries significant clinical and forensic implications: an average of 257 bites per year are estimated, with only one fatality. This epidemiological context necessitates careful consideration of the standard of care and professional liability The article aims to outline the clinical and medico-legal dimensions of viper bites within both national and international contexts. Methods: We report the case of a 40-year-old woman bitten by a viper in an urban environment. At the initial emergency department visit the presentation was classified as Grade 0 (“dry bite”) according to Boels and, after 21 h, was discharged in good condition. Three days later, she returned with worsening symptoms and CT imaging revealed intrafascial and subcutaneous edema. The subsequent onset of complications prompted a criminal malpractice investigation. Therefore, we performed a PubMed search which yielded 125 records; after applying eligibility criteria, 33 articles were included, supplemented by manual reference checking for a total of 60 sources reviewed. Results: Comparison with the Australian model suggests the need for more standardized care pathways in Italy, while accounting for local toxicological and epidemiological specificities. Conclusions: This case and the accompanying literature analysis highlight that, even in low-incidence settings, structured patient communication, multidisciplinary management, collaboration with Poison Control Centers, and adherence to good clinical practice are crucial for patient safety and for mitigating medico-legal risk.

Forensic Sciences doi: 10.3390/forensicsci6010026

Authors: Federica Spadazzi Dalila Tripi Miriam Ottaviani Paola Frati Mauro Arcangeli Gianpietro Volonnino

Introduction: Gender-based violence and femicide represent the most extreme manifestation of a deep-rooted cultural distortion embedded within patriarchal social structures. In this study, adopting a comparative and multidisciplinary approach, we analyzed the evolution of international legislation and the major historical milestones in the protection of women’s rights and the prevention of gender-based violence at both the global and Italian levels. Specific protocols such as the “Pink code” were examined, with particular attention to medico-legal implications and the clinical management of victims, highlighting how violence against women continues to be fuelled by stereotypes, discrimination, and unequal power relations. Materials and Methods: Gender-based violence and femicide were examined from both national and international perspectives. A total of 73 scientific articles in English and 28 legal sources were selected from an initial pool of 918 publications, through a narrative review with a structured search strategy of international and Italian legislation and scientific literature. Electronic databases (PubMed and Google Scholar) were searched for the period 2000–2025. Only original observational studies, medico-legal analyses, epidemiological reports, and forensic case series were included. Cases primarily related to pregnancy, migration, infanticide, suicide, or substance abuse were excluded to reduce heterogeneity and focus on violence rooted in gender-based power asymmetries. Results: The legislative analysis shows a progressive strengthening of protection mechanisms, particularly between 2012 and 2023, following the ratification of the Istanbul Convention, the increase in intimate partner violence, and the COVID-19 pandemic. In Italy, the repeal of discriminatory norms and the introduction of specific legislative measures have led to increased attention toward prevention, protection, and prosecution of gender-based violence. Protocols such as the ‘Pink Code’, an Italian hospital-based multidisciplinary pathway activated mainly in emergency departments for the early identification, clinical care, medico-legal documentation, and judicial protection of victims of gender-based violence, have improved multidisciplinary management of victims within healthcare and judicial settings, although significant challenges remain regarding the full enforcement of legislation and the effective protection of women. The analysis focuses on female victims, in accordance with the Italian legal definition of gender-based violence, while other forms of gender-related violence were considered beyond the scope of this review. Conclusions: Despite substantial legal advances, combating gender-based violence clearly requires an integrated approach that combines prevention, assistance, and prosecution. Strengthening collaboration among institutions, healthcare services, and the judicial system—consistent with international recommendations—is essential to ensure an effective and rights-based response to victims. Overcoming the cultural and social barriers that perpetuate violence remains a fundamental priority, alongside promoting genuine gender equality.

Forensic Sciences doi: 10.3390/forensicsci6010025

Authors: Maria José Amorim Inês Morais Caldas Daniel Pérez-Mongiovi Alexandra Teixeira

Background/Objectives: When analyzing human skeletal remains for human identification, the assessment of sexual dimorphism is fundamental because it underlies sex estimation, a key parameter of the biological profile, which reduces the number of candidates to approximately one half. Ideally, the most dimorphic bones (the pelvis and long bones) are used, but this analysis may be compromised when the skeletal remains are badly preserved, which is often the case in older skeletons. It is, therefore, necessary to investigate sexual dimorphism in alternative skeletal elements as the quantity and quality of bone structures present in ancient skeletons represent crucial aspects when assessing biological differences between the sexes. The present study aims to evaluate the degree of sexual dimorphism in selected skeletal elements as a preliminary step toward identifying sexually dimorphic parameters with potential applicability in future research on poorly preserved skeletons. Methods: A metric assessment of sexual dimorphism was performed on the clavicle, sternum, and seventh cervical vertebra from a 20th Century collection of identified skeletons from the Portuguese population from CESPU (CEIC), showing a variable degree of bone preservation. Results: Our preliminary data suggest that although all bones analyzed may have exhibited some degree of sexual dimorphism, five parameters—sternal body length, manubrium width, first stern-vertebrae width, clavicle maximum length, and cervical vertebral body height—showed the most promising results. Nevertheless, further studies are needed to validate these observations, using a bigger sample of badly preserved bones. Conclusions: These results represent a preliminary assessment of sexually dimorphic parameters, which may be of interest in circumstances where skeletal elements are poorly preserved.

Forensic Sciences doi: 10.3390/forensicsci6010024

Authors: Santori Francesco Fratini Riccardo Visi Giacomo Romanelli Marta Maiolatesi Federica Leila Fabiani Pasquale Giuseppe Macrì

Background: Listeria monocytogenes is an ubiquitous foodborne pathogen capable of persisting in food-processing environments, domestic settings, and water systems; consequently, the mere co-detection of the microorganism in a patient and in a food product is insufficient to support criminal causation. In judicial proceedings, attribution of invasive listeriosis to a specific food and food business operator requires a pathogen-specific evidentiary chain that exceeds the standards used for public health surveillance. Methods: We performed a retrospective medico-legal and methodological analysis of a fatal listeriosis case that triggered criminal prosecution for foodborne homicide, systematically assessing the investigative chain according to temporal plausibility, food traceability, compliance with European and ISO microbiological standards, interpretation of whole-genome sequencing (WGS) data for an ubiquitous pathogen, and clinical causation of death. Results: The analysis identified critical weaknesses in the accusatory reconstruction, including incompatibility with established incubation periods, lack of proof that the sampled food lot corresponded to the product actually consumed, deviations from standard microbiological procedures, reliance on a poorly documented “clinical” isolate for WGS, non-specific genomic findings within a widely disseminated clonal complex, and the presence of plausible alternative environmental reservoirs. Clinically, the immediate cause of death was more consistent with hypovolemic shock due to uncontrolled gastrointestinal bleeding than with ongoing listerial sepsis. Conclusions: This article demonstrates how, in cases involving ubiquitous pathogens, failure to adopt a pathogen-specific investigative and interpretative framework may lead to methodologically fragile criminal allegations, potentially increasing litigation burden and costs for the justice system. A methodologically robust approach integrating microbiology, genomics, epidemiology and medico-legal causation analysis is essential for fair and scientifically sound criminal proceedings.

Forensic Sciences doi: 10.3390/forensicsci6010023

Authors: Tooba Siddiqui Peter Zioupos Nicholas Marquez-Grant

Background: Stature estimation is a key parameter in biological profiling of human skeletal remains and thus in aiding in the identification process of the deceased. Various methods, including anatomical approaches, multifactorial regressions and organic correlation approaches, have been developed to estimate stature. Challenges arise from incomplete or altered remains, outdated reference samples, and the lack of population specific studies. This paper critically examines genetic influences and population-specific factors, the present status quo, recent developments and the challenges in stature estimation in forensic anthropology. Methods: The study appraises the current status, challenges, and future directions of stature estimation in forensic anthropology and bioarchaeology. The open-source literature is systematically identified, and relevant osteological and imaging-based studies are screened and prioritised when they report original empirical data or clearly defined methodological developments, enabling consistent extraction of sample and population descriptors, modelling strategies, and limitations. Included studies are then synthesised comparatively and summarised in a study table to support direct cross-study evaluation. Results: Stature estimation accuracy is shown to depend on population, ancestry, sex, time period, and environment, with cross-population or cross-temporal equation transfer identified as a key source of bias. Limitations include fragmented/altered remains and non-representative reference samples, while newer imaging and probabilistic approaches improve performance when supported by suitable data and explicit error quantification. Conclusions: In the present article, stature estimation is positioned as a context-dependent, evidence-weighted process rather than a fixed calculation.

Forensic Sciences doi: 10.3390/forensicsci6010022

Authors: Othmane Essoubaiy Adnane Hakem Faiza Chbel Hakima Yahia Hicham EL Ossmani Taoufiq Fechtali Brahim El Houate

Background/Objectives: The Casablanca-Settat region of Morocco, located at the interface between Arab and Amazigh cultural zones, has only recently been investigated using autosomal short tandem repeat (STR) markers. The objective of this study was to characterize the genetic diversity and forensic efficiency of 15 autosomal STR loci in the Casablanca-Settat population and to evaluate its genetic relationships with other Moroccan populations. Methods: Fifteen autosomal STR loci were genotyped in 138 unrelated Arabic-speaking individuals from the Casablanca-Settat region. Allele frequencies, Hardy–Weinberg equilibrium, and standard forensic parameters were calculated. The genetic structure of the population was further examined through comparative analyses with 12 previously published Moroccan reference populations using multivariate and phylogenetic approaches. Results: A total of 146 distinct alleles were identified across the 15 loci. D18S51 was the most polymorphic marker (Ho = 0.9203), whereas D3S1358, TPOX, D5S818, and D16S539 exhibited lower allelic diversity. No statistically significant deviation from Hardy–Weinberg equilibrium was detected after correction for multiple testing. The combined power of discrimination exceeded 0.99, and the combined power of exclusion reached 0.99999965, demonstrating the high forensic efficiency of the STR panel. Population structure analyses positioned the Casablanca-Settat population within an intermediate genetic cluster, closely related to central Moroccan populations, consistent with historical gene flow and admixture. Conclusions: This study provides robust autosomal STR reference data for the Casablanca-Settat population, confirming the suitability of these markers for forensic identification in Morocco and offering valuable insights into regional population structure and genetic diversity.

Forensic Sciences doi: 10.3390/forensicsci6010021

Authors: Monica Concato Barbara Di Stefano Raffaella Vetrini Aurora Carnio Maximo Leonardo Franzoni Maria Grazia Calvano Solange Sorçaburu Ciglieri Francesco Introna Paolo Fattorini

Background/Objectives: Personal identification is a cornerstone of Forensic Medicine. Historical records indicated that 27 Italian soldiers were executed by Tito’s army in April 1945 and buried in the mass grave of Ossero (Cres Island, Croatia). Methods: The remains, exhumed in 2019 by Croatian authorities and transferred to the Sacrario d’Oltremare (Bari, Italy), underwent radiographic and anthropological analyses. Genetic analysis was later performed on 147 bone samples, leading to the identification of 10 soldiers. Results: Anthropological analyses revealed commingled remains of at least 34 individuals (15–45 years; 161–181 cm), including eight skeletal elements of female sex. Forensic examination confirmed cranial and post-cranial gunshot wounds. Conclusions: Extensive commingling and fragmentation of the remains prevented full reconstruction of individual skeletons, yet comparison with ante-mortem data supported their identification as the aforementioned soldiers. Unexpected findings, including historically undocumented females, were confirmed by DNA analysis. These findings underscore the value of a multidisciplinary approach to optimize recovery and subsequent forensic and genetic investigations.

Forensic Sciences doi: 10.3390/forensicsci6010020

Authors: Domenico Colloca Matteo Manfredini Fabiano Gentile Alberto Marino Maria Carla Gerra Cristina Dallabona

Background/Objectives: DNA profiling in forensic investigation typically compares genetic profiles, usually derived from the analysis of STR markers. However, this method has limitations when there is no biological reference sample or match in the DNA database. The aim of the current study is thus to replicate, in an Italian cohort, epigenetic markers previously identified in the literature for distinguishing tobacco smokers from non-smokers or estimating chronological age, so as to help narrow down the pool of suspects. Methods: DNA methylation at four CpG dinucleotides located around the cg05575921 site of the AHRR gene, widely associated with tobacco consumption, was measured. Additionally, five CpG dinucleotides in the ELOVL2, FHL2, KLF14, TRIM59, and C1orf132 genes were examined for chronological age estimation in buccal swab samples of 102 volunteers through pyrosequencing. Results: A multiple linear regression model for estimating chronological age shows that ELOVL2-C7, C1orf132-C1, and TRIM59-C7 have a significant effect on age. In this model, the prediction error increases with age. Two logistic regression models were used for determining smoker/non-smoker status, proving that two CpG sites significantly influence the odds of being classified as a smoker. When ex-smokers are included in the non-smoking group, the model correctly classifies the two conditions in about 80% of cases. Conclusions: The results demonstrate that the models generated from pyrosequencing data are useful for identifying tobacco smokers and estimating an individual’s chronological age, particularly for younger subjects. Further studies are needed to develop models with higher predictive accuracy and to integrate these tools into regular forensic practice.

Forensic Sciences doi: 10.3390/forensicsci6010019

Authors: Marion Defontaine Logan Privat Christian Siatka Chloé Scherer Anna Franzoni Michele Rosso Sylvain Hubac Francis Hermitte

Background/Objectives: Rapid DNA systems accelerate STR profiling but often require the consumption of the entire swab, limiting confirmation testing or downstream analyses. We previously validated a simple subsampling protocol for blood swabs on the RapidHITTM ID, using a rigid subungual mini-swab (Copan Italia S.p.A). A new version of this instrument has recently been released, featuring redesigned software and consumables. The RapidINTELTM Plus sample cartridge now enables two distinct lysis/extraction protocols, expanding analytical possibilities for rich biological traces. We evaluated subsampling performance using the subungual mini-swab and microFLOQ® swabs (Copan Italia S.p.A), and assessed feasibility for both blood and buccal reference swabs. Methods: Whole blood from four donors was deposited onto regular Copan swabs (10 µL) or microFLOQ® swabs (1 µL). A comparison was performed between the direct analysis of blood swabs using a RapidHITTM ID V1 (RapidINTELTM cartridge) and a RapidHITTM ID V2 (RapidINTELTM Plus cartridge, GENERAL protocol). Subsequently, both the GENERAL and SPECIALIZED protocols were tested after subsampling from primary blood or buccal swabs dried for 24 h using either a subungual mini-swab or a microFLOQ®. Results: Blood-swab subsampling on the V2 produced usable STR profiles with both the subungual mini-swab and the microFLOQ®. The subungual mini-swab was compatible with both the GENERAL and SPECIALIZED protocols. For blood applications, microFLOQ® fiber treatment showed no inhibitory effects. Reference buccal swabs were successfully analyzed with the RapidINTELTM Plus cartridge, either directly (regular swab) or via subungual subsampling under both protocols. In contrast, in this feasibility dataset (single analysis per donor per condition), subsampling a reference swab with microFLOQ® did not yield suitable profiles for RapidINTELTM Plus analysis under the tested conditions. Conclusions: This feasibility study indicates that the subsampling strategy can be applied on the RapidHITTM ID V2, particularly using subungual mini-swabs, to retain the primary swab for potential downstream testing while maintaining usable STR profile quality for blood and buccal reference workflows under the tested conditions.

Forensic Sciences doi: 10.3390/forensicsci6010018

Authors: Persefoni Lambrou Eleftherios G. Kaklamanos Neophytos Christodoulou Kostis Giannakopoulos

This review explores the vital role of orthodontic specialty and orthodontic records in forensic investigations, emphasizing their contribution to human identification. It synthesizes current literature on methodologies, advantages, limitations, and future perspectives, positioning orthodontics as a valuable adjunct within the forensic toolkit. Collaboration between orthodontists, forensic scientists, and AI technologies will further strengthen the reliability and speed of human identification, reinforcing justice and humanitarian efforts worldwide.

Forensic Sciences doi: 10.3390/forensicsci6010017

Authors: Lilioara-Alexandra Oprinca-Muja Adrian-Nicolae Cristian George-Călin Oprinca Elena Topîrcean Alina Cristian Manuela Mihalache Cosmin Mihalache Marius Florentin Popa Silviu Morar

Burnout syndrome is increasingly recognized as a significant occupational health issue, characterized by emotional exhaustion, depersonalization, and a reduced sense of personal accomplishment. It predominantly arises from chronic work-related stress, but recent research has highlighted the role of genetic and epigenetic factors in determining individual vulnerability to burnout. This review aims to synthesize findings regarding the genetic footprints of burnout, focusing on genes related to stress regulation, including the 5-HTT (serotonin transporter) gene, BDNF (brain-derived neurotrophic factor) gene, and NR3C1 (glucocorticoid receptor) gene. Twin studies reveal that burnout is moderately heritable, with genetic factors accounting for 33–36% of the variability in burnout-related traits, such as emotional exhaustion and performance-based self-esteem. However, burnout risk seems highly driven by non-shared environmental factors, such as work stress, lack of social support, and personal coping mechanisms. Specific genetic polymorphisms in the serotonergic system (5-HTT) and HPA axis genes (NR3C1, FKBP5) have been linked to increased burnout susceptibility, particularly in individuals exposed to chronic job strain or early-life stressful situations. Variations in 5-HTT rs6354 and HTR2A rs6313 are associated with altered stress reactivity, while polymorphisms in NR3C1 and FKBP5 contribute to dysregulation of the HPA axis, which influences cortisol secretion patterns in response to stress. Increased methylation in genes like BDNF and SLC6A4 has been observed in individuals with burnout, suggesting that environmental stressors may lead to lasting changes in gene expression, contributing to the syndrome’s development. Studies on telomere length have shown that burnout is associated with accelerated cellular aging, with individuals exhibiting shorter telomeres, particularly during high-stress periods. These findings hold particular relevance for professionals within the forensic and justice systems, including law enforcement, the judiciary, and forensic experts, who operate under chronic, high-stakes stress. We examine how understanding the biological basis of burnout can inform more objective ‘fitness-for-duty’ evaluations and provide a scientific framework for distinguishing physiological exhaustion from professional negligence in legal contexts.

Forensic Sciences doi: 10.3390/forensicsci6010016

Authors: Roman Kuruc Andrea Szórádová Jozef Šidlo Michaela Neszméry Ľuboš Nižnanský

Background/Objectives: Hanging is the most common method of suicide in most countries worldwide. It is characterized by high lethality, technical simplicity, and typical autopsy findings. Autopsy plays a crucial role in determining the cause and mechanism of death. While external injuries are relatively consistent, internal findings show considerable variability in the literature. The aim of this prospective study was to analyze 660 cases of suicidal hanging over a ten-year period, focusing on the occurrence of forensically relevant internal autopsy findings. Methods: The study was conducted at the Department of Forensic Medicine in Bratislava between 2015 and 2024. All cases underwent standardized complete autopsy, including histology, toxicology, and analysis of death circumstances. Recently reported thoracic aortic adventitial hemorrhages described in 2024 were evaluated only in a targeted subset of cases examined between July and December 2024. Statistical evaluation was performed using the chi-square test to identify associations between internal findings and suspension type, knot location, age, sex, and body weight. Results: The argent line was present in 61.1% of cases, most frequently with posterior knot placement and complete suspension. Neck muscle hemorrhages occurred in 53.8%, predominantly at the periosteal-clavicular attachment of the sternocleidomastoid muscle, with higher incidence in complete and anterior suspension. Amussat’s sign was observed in 10.2% of cases, and Etienne-Martin’s sign in 1.1%. Fractures of the laryngo-hyoid complex were present in 49.7%, mainly in cases with complete suspension and posterior knot location. Cervical spine injuries were detected in 2.6%, predominantly in older males and with anterior knot placement. Simon’s hemorrhages occurred in 35.2%, mainly in younger individuals and complete suspension. Hemorrhages in the intestinal wall were detected in 7.4%, and rectal hemorrhages in 1.1% of cases. In the targeted 2024 subset, no thoracic aortic adventitial hemorrhages were identified. Conclusions: The findings suggest the forensic relevance of several internal findings associated with hanging, while emphasizing that the results were obtained using a uniform and consistently applied autopsy protocol. They also indicate the need for further research, particularly regarding recently reported adventitial hemorrhages of the thoracic aorta, which were assessed only in a limited subset of cases during the final months of the study and were not identified in our material.

Forensic Sciences doi: 10.3390/forensicsci6010015

Authors: Antonio Della Valle Eleonora Lo Muzio Lorenzo Lo Muzio Anna Paola Fabrocini Maria Eleonora Bizzoca

Background and Clinical Significance: Fatal attacks by domestic dogs, particularly against familiar owners, are rare but represent a significant forensic and public health concern. Understanding the dynamics and forensic features distinguishing predatory aggression from postmortem scavenging is essential, especially when cases involve large breeds and vulnerable individuals, such as the elderly. Case Presentation: An 82-year-old man was found dead in his home, presenting extensive mutilation of the head and neck. He lived with his daughter and her four-year-old male German Shepherd. The dog exhibited blood and tissue residues on its mouth, forelimbs, and abdomen. Autopsy findings indicated death due to hemorrhagic shock from deep cervical and facial vessel lacerations. There were no defense wounds or classic bite marks; however, massive excision of musculocutaneous tissue from the neck to the scalp suggested active predation rather than postmortem scavenging. The facial and cervical soft tissues were completely avulsed, exposing deep anatomical structures and causing the loss of the right eye and dental elements. No signs of third-party involvement, intrusion, or external aggression were identified, and the dog appeared healthy and unrestrained. Conclusions: The findings support the theory of a fatal attack by the household German Shepherd occurring shortly after the victim’s meal. This case underscores the potential for lethal aggression in domestic dogs toward familiar humans, even in the absence of provocation or prior behavioral concerns. It highlights the importance of awareness and preventive measures when managing large, powerful dog breeds in environments with elderly or otherwise vulnerable individuals.

Forensic Sciences doi: 10.3390/forensicsci6010014

Authors: Muhammad Khairie Basjuri Anita Zara Weinheimer Muhammad Reza Amin Reza Adnan Eddy Saputra Rohmatul Amin Muhammad Jefri Mohd Yusof

Background/Objectives: Nasal morphology is an important craniofacial feature in forensic anthropology for examining population-level variation and informing biological profiling in living individuals. Despite genetic, historical, and cultural similarities between Malaysian and Indonesian populations, population-specific nasal anthropometric data remain limited. This study integrates nasal anthropometry with multivariate analytical approaches to examine population- and sex-related variation in nasal morphology among Malaysian and Indonesian adults. Methods: A cross-sectional anthropometric study was conducted among Malaysian and Indonesian adults. Nasal breadth and nasal height were measured using standardised techniques, and the nasal index was calculated. Univariate analyses were performed to assess population- and sex-related differences. Multivariate analyses, including principal component analysis (PCA), hierarchical cluster analysis (HCA), and partial least squares (PLS), were applied to eleven nasal parameters and the nasal index to evaluate patterns of population differentiation. Results: Malaysian participants predominantly exhibited leptorrhine nasal types, whereas mesorrhine nasal types were more prevalent among Indonesian participants. Significant differences were observed in nasal height and nasal index between populations, while nasal breadth did not differ significantly. Sex-related variation was evident, with males generally exhibiting larger nasal dimensions and higher nasal indices than females. Multivariate analyses demonstrated clear population separation, with modelling indicating that population group contributed more strongly to morphological differentiation than sex. Conclusions: The findings provide population-specific soft-tissue nasal data for Malaysian and Indonesian groups and support the use of multivariate analytical frameworks in forensic anthropological research.

Forensic Sciences doi: 10.3390/forensicsci6010013

Authors: Alessia Leggio Ricardo Ortega-Ruiz Giulia Iacobellis

The application of advanced imaging techniques, particularly computed tomography (CT), photogrammetric scanning, and three-dimensional reconstructions of body surfaces and skeletal remains, is becoming a crucial component of Forensic Anthropology. These tools enable a non-invasive and highly standardized analysis of both intact cadavers and human remains recovered from terrestrial or aquatic environments, providing reliable support in identification processes, traumatological reconstruction, and the assessment of taphonomic processes. In the context of estimating the Post-Mortem Interval (PMI) and the Post-Mortem Submersion Interval (PMSI), digital imaging allows for the objective and reproducible documentation of morphological changes associated with decomposition, saponification, skeletonization, and taphonomic patterns specific to the recovery environment. Specifically, CT enables the precise assessment of gas accumulation, transformations in residual soft tissues, and structural bone modifications, while photogrammetry and 3D reconstructions facilitate the longitudinal monitoring of transformative processes in both terrestrial and underwater contexts. These observations enhance the reliability of PMI/PMSI estimates through integrated models that combine morphometric, taphonomic, and environmental data. Beyond PMI/PMSI estimation, imaging techniques play a central role in anthropological bioprofiling, facilitating the estimation of age, sex, and stature, the analysis of dental characteristics, and the evaluation of antemortem or perimortem trauma, including damage caused by terrestrial or fauna. Three-dimensional documentation also provides a permanent, shareable archive suitable for comparative analyses, ensuring transparency and reproducibility in investigations. Although not a complete substitute for traditional autopsy or anthropological examination, imaging serves as an essential complement, particularly in cases where the integrity of remains must be preserved or where environmental conditions hinder the direct handling of osteological material. Future directions include the development of AI-based predictive models for PMI/PMSI estimation using automated analysis of post-mortem changes, greater standardization of imaging protocols for aquatic remains, and the use of digital sensors and multimodal techniques to characterize microstructural alterations not detectable by the naked eye. The integration of high-resolution imaging and advanced analytical algorithms promises to further enhance the reconstructive accuracy and interpretative capacity of Forensic Anthropology.

Forensic Sciences doi: 10.3390/forensicsci6010012

Authors: Gayathri Narayanan Prabhadevi Muhammed Siddik Abdul Samad Ayona Jayadev Deepa Indira Nair Gevargis Muramthookil Thomas Geena Prasad

Emerging contaminants (ECs) are trace-level chemical and biological compounds detected in the environment, particularly in water, including personal care products, microplastics, nanoplastics, antibiotic resistance genes, etc., which have the potential to endanger ecosystems and human health. Forensic techniques, which are used to detect and trace the sources of ECs with specificity and accuracy, like gas and liquid chromatography coupled with mass spectrometry, Fourier transform infrared spectroscopy, inductively coupled plasma mass spectrometry, isotope ratio mass spectrometry, remote sensing, geographical information systems, and statistical and machine learning approaches are discussed in this study. The present analysis also outlines the types of emerging contaminants, along with their possible sources, including domestic and municipal wastewater, wastewater from healthcare institutions, urban runoff and stormwater, industrial and commercial discharges, effluents from agricultural and animal husbandry, and pollution from recreational activities in water. The review concludes by emphasizing the future need for real-time detection systems, integrated data analysis, and stronger regulatory frameworks to manage ECs more effectively.

Forensic Sciences doi: 10.3390/forensicsci6010011

Authors: Suthisa Leasen Panida Lorwongtragool Sittichoke Chaiwan Montri Donphoongpri

Background: In forensic ballistics, identifying ammunition types on physical evidence is critical, particularly when metallic residues are minimal. This study comparatively analyzes the elemental signatures deposited by two common projectiles—Full Metal Jacket (FMJ) (Cu/Zn jacket) and Lead Round Nose (LRN) (exposed Pb core)—on complex targets, including pig bone/tissue and mango wood. Methods: Using a semi-automatic handgun at an intermediate range of 5.0 m, residues were examined through high-resolution benchtop Micro-XRF (M4 Tornado) for micro-spatial analysis and Portable XRF (Elio) for rapid field characterization. Additionally, fresh pork leg samples were subjected to a 3-month environmental degradation period to assess trace persistence. Results: Observations indicated that LRN projectiles exhibit markedly elevated Lead (Pb) concentrations along the wound track in bone, hence confirming Pb as a reliable indicator for unjacketed ammunition; specifically, the median Pb concentrations at bullet wiping were 10.39 wt% for M4 and 7.34 wt% for Elio. Conversely, FMJ traces remain strictly confined to the surface bullet wipe area, with median concentrations of Pb, Cu, and Zn being 2.21 wt%, 0.24 wt%, and 0.59 wt% via M4, respectively. Statistical analysis showed a strong correlation for high-concentration elements on tissue, but significantly greater variance on wooden surfaces where FMJ traces exhibited a very weak negative correlation (r = −0.2774) due to minimal and irregular metal transfer. Taphonomic evaluation revealed that the Pb signature from LRN is exceptionally stable (r ≈ 0.9999) even after decomposition, while FMJ signatures are highly sensitive to environmental exposure. Conclusions: This research underscores the necessity of high-sensitivity Micro-XRF (M4) for definitive ammunition verification, providing a refined analytical framework for shooting incident reconstruction even involving degraded remains or complex environmental scenes.

Forensic Sciences doi: 10.3390/forensicsci6010010

Authors: Eliana Santos Ruben Maranho Francisco Curate

Background/Objectives: The identification of individuals from human remains is crucial in any scenario where their identity is unknown. The study of ontogenetic allometry, which refers to proportional changes in the shape and size of bones during growth, provides important baseline information for constructing biological profiles. Methods: This study focuses on the analysis of the ontogenetic allometry of the scapula in Portuguese reference skeletal samples, using geometric morphometric techniques. The sample includes 140 individuals (67 females, 73 males), ranging from birth to 89 years old. Scapulae were photographed, and seven landmarks and forty semi-landmarks were digitized using the “tps” programs. Statistical analyses were performed using the MorphoJ (v. 1.08.02) and PAST (v. 5.2) programs. Results: The results point to a significant and continuous growth of the scapula in the early stages of life, with a tendency to stabilize after adolescence. Centroid size significantly influenced shape variation across the full sample. Conclusions: These findings provide a descriptive baseline of scapular development that can aid future anthropological and forensic research, including studies on population variation and age-related morphological trajectories.

Forensic Sciences doi: 10.3390/forensicsci6010009

Authors: Leandro Oka-Duarte Bruno Ferreira Marcelo Firmino de Oliveira

Background/Objectives: MDMA and MDA are among the stimulant drugs most frequently encountered in forensic casework, and oral fluid represents a practical biological matrix for their detection. However, liquid oral fluid requires refrigeration, is susceptible to degradation, and can be logistically demanding for routine laboratories. Dried Oral Fluid Spots (DOFS) offer a low-cost and stable alternative. This study aimed to develop and validate a DOFS-based analytical workflow for quantifying MDMA and MDA using liquid chromatography and a diode-array detector. Methods: Watercolor paper was selected as the substrate and pretreated with diluted nitric acid to improve analyte desorption. DOFS were prepared using 150 µL of pooled oral fluid, dried for 4 h, and extracted with methanol. Chromatographic separation was performed on a phenyl column using aqueous TFA and acetonitrile mobile phase. Method validation followed the ICH M10 criteria. Results: The method showed linear responses between 12.5 and 5000 ng mL−1, with LOD and LLOQ of 6 and 12 ng mL−1 for both analytes, respectively. Precision and accuracy met acceptance criteria across all QC levels. Recoveries ranged from 84% to98%. DOFS samples demonstrated adequate stability under multiple storage and handling conditions. Conclusions: The optimized DOFS–LC–DAD workflow offers a robust, low-cost, and flexible approach for the analysis of MDMA and MDA in oral fluid for laboratory-based or semi-controlled collection environments. Its compatibility with both LC- and GC-based detectors enhances applicability in diverse forensic laboratory settings.

Forensic Sciences doi: 10.3390/forensicsci6010008

Authors: Antony Cevallos

Forensic anthropology, a specialized branch of biological anthropology, plays a crucial role in the identification of human remains, particularly when conventional methods such as fingerprinting are not applicable. In Ecuador, its relevance has increased in response to challenges such as intentional deaths, forced disappearances, violence, mass fatalities, and migration-related deaths. Despite its growing importance, the field faces significant limitations, including restricted access to advanced technologies, limited training opportunities for local forensic anthropologists, and insufficient resources for research and the application of advanced methodologies for victim identification. This article examines the development and current state of forensic anthropology in Ecuador, emphasizing the urgent need for population-specific standards, the establishment of a national osteological collection, and stronger institutional support. It also highlights the contributions of bioarchaeological research and its potential to enhance forensic practices. By analyzing the challenges of identifying skeletonized human remains and other instances of human rights violations, the study underscores the necessity of advancing forensic anthropology in the country. The article further discusses how interdisciplinary efforts have contributed to forensic knowledge in Ecuador and concludes by emphasizing the importance of ethical guidelines, technological integration, and improved infrastructure to strengthen forensic anthropology as both a scientific discipline and a humanitarian tool.

Forensic Sciences doi: 10.3390/forensicsci6010007

Authors: Matteo Antonio Sacco Lucia Tarda Saverio Gualtieri Maria Cristina Verrina Isabella Aquila

Background: Cigarette burns represent a well-established forensic indicator of inflicted injury, frequently encountered in cases of domestic violence. Clinical significance: Their morphological consistency and anatomical distribution offer valuable elements for differentiating between intentional and accidental trauma. Case Presentation: In this study, we report the case of a 40-year-old woman who presented with multiple cutaneous lesions attributed to repeated assaults by her intimate partner. The forensic medical examination revealed five discrete scars characterized by sharply demarcated borders, circular to oval shapes, and dimensions ranging from 0.7 to 1.5 cm. These lesions were anatomically located in regions not typically accessible for self-infliction. To reinforce the diagnostic interpretation and assess reproducibility, a controlled experimental protocol was conducted using porcine skin matrices. Cigarette burns were recreated under variable conditions of contact pressure and exposure duration. The lesions produced on the biological substrate exhibited morphological features consistent with those observed in the patient, suggesting compatibility with cigarette-induced thermal injury. Conclusions: These findings provide circumstantial support for the forensic interpretation but must be considered within the limitations of the experimental model. This integrated approach underscores the relevance of combining clinical forensic documentation with experimental validation to support medico-legal conclusions in cases of suspected interpersonal violence.

Forensic Sciences doi: 10.3390/forensicsci6010006

Authors: Csongor Herke

Background/Objectives: This systematic review examines how artificial intelligence (AI) is transforming fingerprint and latent print identification in criminal investigations, tracing the evolution from traditional dactyloscopy to Automated Fingerprint Identification Systems (AFISs) and AI-enhanced biometric pipelines. Methods: Following PRISMA 2020 guidelines, we conducted a literature search in the Scopus, Web of Science, PubMed/MEDLINE, and legal databases for the period 2000–2025, using multi-step Boolean search strings targeting AI-based fingerprint identification; 68,195 records were identified, of which 61 peer-reviewed studies met predefined inclusion criteria and were included in the qualitative synthesis (no meta-analysis). Results: Across the included studies, AI-enhanced AFIS solutions frequently demonstrated improvements in speed and scalability and, in several controlled benchmarks, improved matching performance on low-quality or partial fingerprints, although the results varied depending on datasets, evaluation protocols, and operational contexts. They also showed a potential to reduce certain forms of examiner-related contextual bias, while remaining susceptible to dataset- and model-induced biases. Conclusions: The evidence indicates that hybrid human–AI workflows—where expert examiners retain decision making authority but use AI for candidate filtering, image enhancement, and data structuring—currently offer the most reliable model, and emerging developments such as multimodal biometric fusion, edge computing, and quantum machine learning may contribute to making AI-based fingerprint identification an increasingly important component of law enforcement practice, provided that robust regulation, continuous validation, and transparent governance are ensured.

Forensic Sciences doi: 10.3390/forensicsci6010005

Authors: Francesco Gabrielli Francesco Calabrò Lorenzo Franceschetti Silvio Chericoni Valentina Bugelli

Background. Determining the cause and manner of death in scenes involving multiple and putrified bodies found in the same environment is a real challenge for forensic pathologists. While common scenarios include fires, vehicle crashes, and natural disasters, one of the most common causes is drug intoxication or poisoning, and the scene must be carefully evaluated based on circumstantial evidence. Carbon monoxide (CO) (also called “the silent killer”) remains one of the leading agents capable of producing simultaneous fatalities. In multi-body scenes, distinguishing between homicide–suicide, double suicide, and accidental deaths adds further complexity. The aim of this study is to highlight the limitations of toxicological and pathological investigations in advanced putrefaction and to emphasize the role of scene investigation in the interpretation of suspected CO-related deaths. Methods. The authors report a case of suspected CO intoxication involving two bodies in an advanced stage of decomposition recovered from the same room. The scene investigation, coupled with the presence of a malfunctioning combustion source, raised suspicion of CO exposure; however, analytical interpretation was severely constrained by the altered condition of biological samples. Results. Advanced decomposition magnifies these challenges. Putrefactive changes can mimic traumatic injuries, hide hypostasis, and compromise both macroscopic and microscopic evaluations due to autolysis and gas formation. Toxicological investigations are frequently hindered by the degradation or absence of key biological matrices such as blood, cavity fluids, or vitreous humor, rendering carboxyhaemoglobin quantification unreliable or impossible. These limitations may lead to incorrect medico-legal conclusions. Conclusions. Determining the cause and manner of death in complex multi-body scenes requires careful evaluation of circumstantial evidence and scene investigation, particularly when advanced decomposition compromises biological analyses and toxicological interpretation.

Forensic Sciences doi: 10.3390/forensicsci6010004

Authors: Myrsini Voulgari Ioanna Anastopoulou Konstantinos Moraitis

Background/Objectives: A common challenge in both forensic and bioarchaeological research is commingling, the intermixing of skeletal material originating from multiple individuals or contexts. To tackle that problem past reassociation methods primarily relied on visual assessment or metric comparisons. However, recent advances in geometric morphometrics show strong potential for improving the sorting of commingled remains. This study applies a three-dimensional (3D) geometric morphometric method to evaluate its effectiveness in reassociating adjoining thoracic vertebrae. Methods: Two vertebral pairs, T4–T5 and T5–T6, from 65 and 73 individuals, respectively, were analyzed. These pairs were chosen due to limited anatomical variability, while they were also the most consistently preserved pairs. All specimens were scanned using a structured-light 3D scanner, and the dataset was derived from three Greek skeletal collections representing different geo-chronological contexts. Fourteen anatomical landmarks were placed on the superior rim and articular facets of the lower vertebra and mirrored onto the lower rim and facets of the adjoining upper vertebra. To remove the size effects the landmark coordinates were converted to Procrustes coordinates, while examining morphological similarity was quantified using Euclidean distances. For each pair, the vertebrae with the smallest Euclidean distances were considered the most probable true anatomical matches. Results: The correct T4–T5 match fell within the three smallest distances in 66.2% of cases, while for the T5–T6 pair, correct matches were found between the first three possible matches in a percentage of 43.8%. These findings indicate that the method can eliminate roughly 50–70% of incorrect matches and therefore narrow the plausible pairings. Conclusions: Future research incorporating more pairs and an expanded landmark dataset may result in greater accuracy for reassociation with 3D geometric morphometrics.

Forensic Sciences doi: 10.3390/forensicsci6010003

Authors: Tanja Visković Marija Definis Livia Sliskovic

Background: Postmortem DNA identification of highly decomposed human remains is often limited by the availability and quality of conventional biological samples. Keratinized tissues, such as fingernails, represent a potentially valuable alternative due to their anatomical resistance to environmental degradation, however, their use as primary biological material for DNA profiling remains underreported in forensic practice. Case presentation: We report a case involving the recovery of a highly decomposed body of a missing woman, in which DNA samples were collected from a fingernail and a tooth. DNA extraction was performed using the PrepFiler Forensic DNA Extraction Kit for the fingernail sample and PrepFiler BTA Forensic DNA Extraction Kit for the tooth sample. No usable DNA profile was obtained from the tooth sample; however, the fingernail sample yielded a complete and high-quality STR profile with successful amplification across all 24 loci (GlobalFiler PCR Amplification Kit). Reference buccal swabs from the presumed biological parents were collected for subsequent kinship analysis. Discussion: Kinship analysis based on allele frequencies in the Croatian population resulted in a combined paternity index (CPI) corresponding to a probability of paternity of 99.99999812%, providing strong genetic support for the proposed identity of the deceased. Notably, this is the first documented forensic case in Croatia in which nail material served as the primary—and ultimately successful—biological sample for postmortem identification. Conclusions: This case highlights the evidentiary value of fingernails as a robust, accessible, and forensically valid DNA source in postmortem identification, particularly in cases of advanced decomposition where conventional biological materials are unavailable or degraded. Further studies involving larger sample sets and diverse postmortem conditions are needed to support the broader implementation of nail material in routine forensic identification workflows, particularly within the Croatian medico-legal context.

Forensic Sciences doi: 10.3390/forensicsci6010002

Authors: Matteo Antonio Sacco Saverio Gualtieri Aurora Princi Isabella Aquila

Background/Objectives: Snakebite envenoming remains a critical yet frequently under-recognized cause of mortality in many parts of the world, particularly in tropical and rural areas where access to timely medical care and accurate post-mortem investigation is limited. While clinical and epidemiological data on snakebites have been extensively studied, the forensic characterization of fatal envenomations remains fragmentary and inconsistently documented. This review aims to synthesize the existing literature on autopsy-confirmed snakebite deaths, focusing on the pathological and toxicological evidence that supports cause-of-death determinations in forensic settings. Methods: A comprehensive search of the PubMed NCBI databases identified nine relevant studies, including case reports, retrospective analyses, and systematic reviews. Results: Across these reports, a range of lethal mechanisms were identified, including venom-induced consumption coagulopathy (VICC), acute renal failure (frequently in the setting of rhabdomyolysis and acute tubular necrosis), neurotoxic respiratory arrest, multi-organ necrosis, and myocardial infarction. Histological findings frequently revealed glomerular and tubular necrosis, pulmonary edema and/or hemorrhage, pituitary and adrenal hemorrhage, and cerebral ischemic changes. Toxicological confirmation was achieved in several cases using ELISA and liquid chromatography–mass spectrometry (LC–MS/MS), underscoring the importance of biochemical validation in post-mortem diagnosis and the value of analytical tools beyond ELISA (e.g., immunoaffinity LC–MS/MS, venom-specific immunohistochemistry, zymography for SVMP activity). Conclusions: Our findings highlight the variability in venom effects across snake families—particularly Viperidae, Elapidae, and Lamprophiidae/Atractaspididae—and emphasize the indispensable role of forensic autopsy in distinguishing snakebite envenoming from other causes of sudden or unexplained death. However, significant limitations persist, including inconsistent autopsy protocols, lack of species-specific venom assays, and poor integration of toxicological methods in routine forensic practice. Addressing these gaps through standardized forensic guidelines and improved access to diagnostic tools is essential for enhancing the accuracy of death investigations in envenoming-endemic regions.

Forensic Sciences doi: 10.3390/forensicsci6010001

Authors: Phetcharat Phetnui Chanasorn Poodendaen Narawadee Choompoo Kaemisa Srisen Sitthichai Iamsaard Arada Chaiyamoon Supatcharee Arun Thewarid Berkban Suthat Duangchit

Background/Objective: Accurate biological profile estimation from skeletal remains is fundamental to forensic identification. While the humerus demonstrates considerable sexual dimorphism, population-specific validation data for Thai populations remain limited. This study aimed to develop and independently validate population-specific equations for sex and stature estimation from humeral measurements in Northeastern Thai populations. Methods: This cross-sectional study examined 300 adult humeri (150 male, 150 female) from the Khon Kaen University skeletal collection. Four osteometric measurements (maximum length, midshaft circumference, epicondylar breadth, superior–inferior head diameter) and weight were recorded. The sample was randomly divided into development (n = 200) and validation (n = 100) datasets. Logistic regression for sex estimation and linear regression for stature estimation were developed using stepwise selection. Results: Sex classification achieved 93.5% accuracy in development and 93.0% in independent validation. The optimal model incorporated midshaft circumference, superior–inferior head diameter, and weight, with an area under the curve of 0.977 (95% CI: 0.953–1.000), sensitivity 90.0%, specificity 96.0%, and Cohen’s kappa 0.86. Stature estimation demonstrated a correlation coefficient of 0.81 with a mean absolute error of 4.36 cm (2.74% of the mean stature). Independent validation confirmed minimal performance deterioration for both models. Conclusions: These independently validated, population-specific equations provide accurate and reliable methods for biological profile estimation in Northeastern Thai forensic contexts. The rigorous validation framework supports confident operational application and provides a methodological model for developing regional forensic standards.

Forensic Sciences doi: 10.3390/forensicsci5040080

Authors: Camilla Cecannecchia Elena Giacani Benedetta Baldari Antonello Bellomo Luigi Cipolloni Andrea Cioffi

Introduction: Paternal filicide is a rare and complex form of intrafamilial homicide, frequently associated with underlying psychopathology, interpersonal conflict, and psychosocial stressors. While maternal filicide has been more extensively studied, cases involving fathers—especially those employing multiple homicidal methods—remain significantly underrepresented in the forensic literature. This paper presents an unusual case of paternal filicide involving combined lethal methods, contextualized through a narrative review of comparable cases. Methods: A comprehensive forensic-pathological and psychiatric investigation was conducted following the homicide of an 8-year-old boy, killed by his father through a combination of asphyxiation and stabbing. A narrative literature review was performed using PubMed, Scopus, and Google Scholar, focusing on case reports and case series concerning paternal filicide. Particular attention was paid to homicidal methods, motivational dynamics, psychiatric comorbidities, and post-crime behavior. Results: The child’s body was found concealed in a building, in a bed storage drawer, with packing tape tightly wrapped around the mouth and nose and a kitchen knife embedded in the neck. No defensive wounds were observed, suggesting a sudden and unopposed assault, likely facilitated by the victim’s trust in the perpetrator. Autopsy findings revealed signs of asphyxiation and three stab wounds to the chin, neck, and thorax, involving vital structures such as the thyroid cartilage and heart. The father was found in a state of acute alcohol intoxication and subsequently convicted of intentional homicide. The motive appeared to be revenge-related, stemming from a highly conflictual marital separation. The literature review confirmed the predominance of retaliatory motives, frequent substance use, and post-crime suicidal behavior. However, the use of combined homicidal methods and the concealment of the body were found to be exceedingly rare. Conclusions: This case, combined with the literature review, highlights the need for deeper scientific exploration of paternal filicide. Comprehensive forensic and psychiatric assessments are essential to identify recurring situational patterns, motivational profiles, sociocultural contexts, and psychiatric vulnerabilities. These findings are critical not only for post-crime evaluations but also for the development of interdisciplinary prevention strategies targeting early warning signs and high-risk family dynamics.

Forensic Sciences doi: 10.3390/forensicsci5040079

Authors: Abhaya Gupta Mridu Sharma Varsha Dabas Kavita Kumari Sameer Saharan

Background/Objectives: The recovery of latent fingerprints from submerged evidence remains a critical challenge in forensic science, as ridge details deteriorate rapidly once under water. This study aims to compare the effectiveness of three established fingerprint development techniques—cyanoacrylate fuming, small particle reagent (SPR), and powder dusting—on non-porous substrates (glass slides and stainless steel blades) immersed in different water types representative of Rajasthan’s Shekhawati region. The objective was to evaluate the influence of water composition and immersion duration on the quality and reproducibility of developed prints. Methods: Experiments were conducted under controlled laboratory conditions. Fingerprints were submerged in hard water, mineral water, and rainwater for durations of 10 min, 1 day, 5 days, and 10 days. Each condition was replicated three times. Developed fingerprints were assessed for ridge clarity using a five-point scoring scale, and the results were statistically analyzed using Chi-Square and correlation tests. Results: Cyanoacrylate fuming consistently produced the highest quality ridge detail across all submersion periods, particularly in mineral and rainwater environments. SPR exhibited moderate effectiveness, while powder dusting showed limited performance under all conditions. Statistical analysis indicated that fingerprint quality was significantly affected by water composition, substrate type, and immersion duration (p < 0.001). Conclusions: The study highlights that fingerprint recovery from submerged non-porous evidence depends strongly on water chemistry and exposure time. Cyanoacrylate fuming is confirmed as the most reliable method, while environmental variables such as ion content and water hardness play decisive roles in fingerprint preservation and visualization.

Forensic Sciences doi: 10.3390/forensicsci5040078

Authors: Woranan Kirisattayakul Gitsadaporn Boonruengudom Phanusit Chalardpongpun Suthat Duangchit Chanasorn Poodendaen Phatthiraporn Aorachon Sitthichai Iamsaard

Background/Objectives: The external occipital protuberance (EOP) is an anatomical landmark with radiological and anthropological implications. Although the morphology and prevalence of EOP have been studied in many populations, data remain lacking for Northeastern Thais. Population-specific characterization of EOP variation may improve diagnostic and forensic accuracy applications. Materials and Methods: This study has investigated the prevalence and morphometry of EOPs using two primary sources: CT brain scans from 750 adult patients (375 males, 375 females) and anatomical investigations of 1060 dry skulls. EOPs were classified as Type I (flat), Type II (crest), or Type III (spur). Measurements for Type II (crest-shaped) EOPs were performed using standardized linear and angular parameters. Data differences were analyzed by sex and age group; intra- and inter-observer reliability was calculated for imaging measurements. Results: The study showed that Type II EOP was most common in both CT (56.1%) and dry skull (64.6%) samples. Type I was significantly more frequent in females (CT: 37.0%; dry skull: 32.8%), while Type III prevalence was higher in males (CT: 28.5%; dry skull: 18.4%). After age 60, the incidence of Type III declined in both datasets. Type II EOPs were significantly larger in males (mean crest length in CT: males 7.1 ± 0.1 mm, females 5.6 ± 0.1 mm; p < 0.001), with notable sex- and age-associated variation in associated angular dimensions. Conclusions: These findings established the first region-specific morphometric reference database for EOP in Northeastern Thais. The demonstrated sexual dimorphism in Type II EOP measurements provided the foundational data that may support future applications in clinical assessment, radiological interpretation, and forensic sex estimation in this population.

Forensic Sciences doi: 10.3390/forensicsci5040077

Authors: Silvina Marques Moura Áurea Madureira-Carvalho Álvaro Azevedo Inês Morais Caldas

Background/Objectives: A global approach to age estimation is being refined to integrate the contributions of skeletal and dental structures. Teeth represent a unique and durable tool for assessing age, particularly valuable when other biological indicators are unavailable. However, age estimation becomes increasingly difficult in older individuals. This study aims to evaluate whether colorimetric analysis of teeth can serve as a reliable and accurate method for estimating chronological age in both living individuals and cadavers. Methods: The study explores the measurement of tooth color as a non-ionizing and non-invasive alternative to conventional age estimation techniques. The approach emphasizes the preservation of forensic evidence and the feasibility of prospective data collection. The methodological framework involves colorimetric assessment supported by emerging technological instruments designed to standardize and improve the objectivity of measurements. Results: Recent advances in color analysis technology have enhanced the precision and reproducibility of dental color measurement. Preliminary findings suggest that age-related color changes in teeth show measurable and consistent patterns, supporting their potential use as indicators of chronological age. Conclusions: Tooth color measurement appears to be a promising complementary tool for age estimation in forensic and clinical contexts. Its non-invasive and reproducible nature offers significant advantages, provided that standardized protocols and validated instruments continue to evolve to ensure accuracy and reliability in practical applications.

Forensic Sciences doi: 10.3390/forensicsci5040076

Authors: Matteo Nioi Ernesto d’Aloja

Determining the postmortem interval remains one of the most persistent and fragmented challenges in forensic science. Conventional approaches—thermal, biochemical, molecular, or entomological—capture only isolated fragments of a single physical reality: the irreversible drift of a once-living system toward equilibrium. This Perspective proposes a unifying paradigm in which death is understood as a progressive rise in entropy, encompassing the loss of biological order across thermal, chemical, structural, and ecological domains. Each measurable postmortem variable—temperature decay, metabolite diffusion, macromolecular breakdown, tissue disorganization, and microbial succession—represents a distinct expression of the same universal law. Within this framework, entropy becomes a dimensionless index of disorder that can be normalized and compared across scales, transforming scattered empirical data into a coherent continuum. A Bayesian formulation further integrates these entropic signals according to their temporal reliability, yielding a probabilistic, multidomain equation for PMI estimation. By merging thermodynamics, information theory, and biology, the concept of death as rising entropy offers a comprehensive physical description of the postmortem process and a theoretical foundation for future computational, imaging, and metabolomic models in forensic time analysis.

Forensic Sciences doi: 10.3390/forensicsci5040075

Authors: Richard Idem Somiari Stephen J. Russell John Feeley Stella B. Somiari

Background: The positive identification of a source of tissue as human plays an important role in various contexts. It is particularly important for investigations concerning tissue and organ trafficking, since unequivocal confirmation is required for legal proceedings involving such cases. While deoxyribonucleic (DNA) methods are considered the gold standard for tissue identification, issues such as degraded DNA or the presence of chemical preservatives can hinder performance and positive identification using DNA techniques. Objectives: The aim of this study was to develop a simple method for presumptive identification of human tissue using standard bottom-up proteomics data. Methods: We identified proteins isolated from human kidney, lung and spleen tissues by bottom-up proteomics and database search using Proteome Discoverer and Sequest HT algorithms. The list of identified proteins was sorted based on liquid chromatography (LC)–mass spectrometry (MS) data metrics such as the number of unique peptides used to identify each protein and the % sequence coverage of an identified protein to determine if any parameter would cluster proteins annotated as human in a distinct category. We found that eliminating proteins identified with fewer than two unique peptides and those with less than 5% sequence coverage resulted in a final list where at least half of the remaining proteins are annotated as human. We applied this data filtration process to blinded LC–MS/MS data from 26 previous experiments to assess accuracy. Results: Using bottom-up proteomics data and the filtration rules established, we identified tissue samples (n = 10), including kidney, spleen, lung, formalin-fixed paraffin-embedded uterus, frozen breast tissue, dry blood and dry saliva as human, and tissue (n = 16) from rat, mouse, bovine, and sheep as non-human, resulting in 100% sensitivity and specificity. Conclusions: The results demonstrate that the list of identified proteins following a standard bottom-up proteomics experiment could be filtered and potentially used as a fast and simple method for presumptive human tissue identification.

Forensic Sciences doi: 10.3390/forensicsci5040074

Authors: Federica Amirante Fortunato Pititto Giuseppe Pulin Roberto Bellacicco Elisa Paladini Gerardo Cazzato Biagio Solarino Maricla Marrone

Background: Starvation represents a specific pathological entity characterized by severe nutritional deprivation leading to multi-organ failure. Despite its forensic relevance, a comprehensive synthesis of autopsy findings remains lacking. Methods: This systematic review was conducted in accordance with PRISMA 2020 guidelines. PubMed, Scopus, and Google Scholar were searched from inception to June 2025 using a pre-specified Boolean query. Eligible studies included case reports, case series and cohort investigations reporting post-mortem evidence of starvation or starvation-related malnutrition. Data extracted encompassed demographic, contextual, macroscopic, histological, and ancillary findings. Results: Fourteen studies were included, comprising 20 individual cases and two population-based cohorts (totaling 1647 deaths). Most cases (75%) involved children, predominantly victims of domestic neglect; adults accounted for 25%, mainly due to anorexia nervosa or voluntary fasting. Six cadavers were severely decomposed or mummified. Across studies, consistent autopsy findings included extreme emaciation, near-total loss of subcutaneous and visceral fat, empty gastrointestinal tract, and diffuse organ atrophy, especially of the liver, heart, thymus, and pancreas. Histology revealed hepatic steatosis, myocardial fibrosis, thymic involution and gelatinous transformation of adipose tissue. Ancillary methods (dual-energy X-ray absorptiometry, stable isotope and anthropological analyses) confirmed malnutrition in decomposed or skeletonized remains. Conclusions: This review delineates the morphological and histopathological hallmarks of starvation and suggests the possible diagnostic value of ancillary techniques in advanced decomposition. The predominance of neglect-related pediatric cases underscores starvation as a forensic indicator of social and caregiving failure. Establishing reproducible morphological and histological indicators may improve the consistency of forensic diagnosis and strengthen the evidentiary basis for determining starvation as a cause of death.

Forensic Sciences doi: 10.3390/forensicsci5040073

Authors: Christine Bailey Autumn Lennartz Maggie Klemm Cameron Matheson Carter Unger Rebecca George

Background: Estimating the postmortem interval (PMI) is complicated by extrinsic environmental and intrinsic individual factors. Methods: Improved accuracy may be achieved through a better understanding of desiccation. This study examines moisture loss and desiccation in human remains in western North Carolina, validating previous research in central Texas. Ten donated individuals were placed across three seasonal trials at Western Carolina University’s Forensic Osteology Research Station (FOREST). Soft tissue moisture measurements were recorded from 20 locations on the body using a Delmhorst RDM-3TM meter, and environmental data were recorded on-site. Results: Moisture content declined rapidly until ~500 accumulated degree days (ADD), after which patterns became highly variable. Linear mixed-effects models identified temperature as the strongest predictor of moisture loss, particularly in spring and fall, while precipitation was the most influential in summer, coinciding with rapid skeletonization. Compared to central Texas, western North Carolina exhibited less consistent moisture loss patterns and greater environmental variability. Fixed effects explained 36–63% of moisture variation across body regions, with conditional R2 values modestly higher when accounting for individual differences. Conclusions: These findings highlight the importance of region-specific research for PMI estimation.

Forensic Sciences doi: 10.3390/forensicsci5040072

Authors: Antonieta Zuniga Yolly Molina Karen Amaya Zintia Moya Patricia Soriano Digna Pineda Yessica Pinto Isaac Zablah

Background: The Tawahka ethnic group, with approximately 2690 individuals in northeastern Honduras, represents one of the country’s smallest indigenous communities. No genetic studies have been published on this population, and population-specific databases are essential for forensic applications. Methods: Allele frequencies for 23 autosomal short tandem repeats (STRs) loci were analyzed in 100 unrelated Tawahka individuals (61 females, 39 males) from the municipality of Wampusirpi. Deoxyribonucleic acid (DNA) was extracted from blood on Fast Technology for Analysis of nucleic acids (FTA) cards and amplified using the PowerPlex Fusion 6C System. Statistical parameters were calculated using Genepop v4.2 and Arlequin v5.3.2.2. Results: All loci showed substantial polymorphism with no Hardy–Weinberg equilibrium deviations after Bonferroni correction (α = 0.0022). Expected heterozygosity ranged from 0.4968 to 0.8812. Combined power of discrimination was 99.9999% and combined chance of exclusion was 99.99%. Conclusions: This first genetic characterization of the Tawahka population provides essential reference data for forensic identification, paternity testing, and population genetics studies. The dataset contributes to understanding indigenous Central American genetic diversity and ensures accurate forensic analyses for individuals of Tawahka ancestry following Combined DNA Index System (CODIS) and European Standard Set (ESS) standards.

Forensic Sciences doi: 10.3390/forensicsci5040071

Authors: Alexandra L. Paradis Sean D. Tallman

Background/Objectives: Forensic anthropological analyses and reports include diverse information that may help in establishing identifications, cause or manner of death determinations, taphonomic alterations, or other circumstances involving decedents. However, within forensic anthropology in the United States, there remains no unifying standardization for analyzing cases or writing reports that can be used independent of work environment, education, or case circumstance, which is potentially problematic for a field with 32.4% of practitioners being sole practitioners, as reported here. While some broad “best practice” guidelines exist, they do not provide a significant, nuanced, or unifying set of methodological, terminology, or reporting standards. Methods: In order to explore whether a lack of standardization is problematic in forensic anthropology, a survey was conducted to explore reporting practices, followed standards, and personal experiences regarding overall practices that lead to case reports. Results: Among the 106 respondents, there were multiple areas of agreement, which suggests that unofficial reporting standardization exists within the field. Further, 71.3% agreed that standardization in case reporting is important, and 60.3% supported the adoption of universal standards. However, other areas, such as cognitive bias, terminology, education, and training experiences, varied among respondents. Standardization is important, as 23.2% reported that their laboratory lacks a Standard Operating Procedure; 28.0% reported that their laboratory lacks reporting guidelines; and 24.4% reported that their casework was not peer reviewed. Discussion/Conclusions: The findings from this study suggest that overarching standardization for report writing, terminology, and methodology should be created to provide a baseline reference for all forensic anthropologists, regardless of work context and educational background.

Forensic Sciences doi: 10.3390/forensicsci5040070

Authors: Gennaro Baldino Pietro Tarzia Gabriele Rotter Simona Calabrese Marija Čaplinskienė Elvira Ventura Spagnolo

Background: Fatal hemorrhage from ruptured varicose veins is a rare event that may mimic violent death given the quantity and distribution of blood at the scene. Accurate determination of cause and manner of death requires careful scene investigation, autopsy, histology, and ancillary methods such as bloodstain pattern analysis (BPA). Case presentation: We describe an 80-year-old woman found deceased at home, with evidence of extensive bloodstaining, no significant external trauma except a small laceration over a varicosity, and ultimately a ruptured branch of the small saphenous vein. The cause of death was hemorrhagic shock due to spontaneous varicose vein rupture. Discussion: This case reinforces the importance of integrating death scene findings (including shoe prints, bloodstain distribution), anatomical dissection, histology, and toxicology to exclude non-natural causes. We also review recent similar cases in the literature, noting risk factors, presentation, and common pitfalls. Conclusions: Forensic investigations of massive bleeding in elderly, isolated individuals should include varicose vein rupture in the differential. A multidisciplinary approach is essential to avoid misclassification of manner of death.

Forensic Sciences doi: 10.3390/forensicsci5040069

Authors: Chanasorn Poodendaen Narawadee Choompoo Kaemisa Srisen Supapit Linlad Jetniphat Chalermrerm Worrawit Boonthai Sitthichai Iamsaard Nareelak Tangsrisakda Supatcharee Arun Suthat Duangchit

Background: Sex estimation from skeletal remains is important for forensic identification, but many methodologies focus on complete elements despite high fragmentation rates in operational contexts. The aim of this study was to develop and validate discriminant function equations for sex estimation between complete and fragmented Thai femora. Materials and Methods: A total of 560 adult femora (280 males and 280 females) were used for measurements of eight osteometric variables. Then, discriminant function analysis was applied to complete femora and anatomically isolated segments, including proximal, diaphyseal, and distal, with leave-one-out cross-validation. Results: All measurements showed significant sexual dimorphism, with percentage differences ranging from 6.56% to 42.27%. Complete femur stepwise analysis achieved 90.47% accuracy by using four optimally selected variables, performing comparably to eight-variable models. Isolated segment accuracies varied substantially: proximal segments achieved 89.64% accuracy, differing by only 0.83 percentage points from complete performance; distal segments demonstrated 86.25% accuracy from bicondylar width alone; and diaphyseal segments achieved 80.88%. Combined segment approaches demonstrated synergistic effects approaching complete femur performance. Conclusions: These population-specific equations provided validated methodologies for sex estimation from complete and fragmentary Thai femora. Anatomical region selection still maintained high classification accuracy despite skeletal incompleteness, in line with the fragmentary conditions commonly encountered in forensic and archeological contexts.

Forensic Sciences doi: 10.3390/forensicsci5040068

Authors: Athina Tousia Dimitris Kouzos Konstantinos Katsos Ioannis Ketsekioulafis Ioannis Papoutsis Artemisia Ntona Nikolaos Georgiadis Theodoros N. Sergentanis Chara A. Spiliopoulou Emmanouil I. Sakelliadis

Background/Objectives: Motorcycle accidents have grown to become a significant public health thread worldwide. Most of the victims are described to be males of young age, some of lower socioeconomic status. Throughout the literature, head and spinal injuries are described as the most common injuries, while chest injuries and lower extremity fractures are also frequently described. Moreover, driving under the influence of alcohol has been widely described as a major predisposing factor. We conducted a retrospective cohort study to categorize risk factors, including demographic information and alcohol/psychoactive substance consumption, as well as pre-existing medical conditions. Correspondingly, we identified the most common injury patterns and attempted to establish a connection between time to hospital and survival rates. Methods: Cases of motorcycle-related road traffic accidents (RTAs) examined by the authors during 2021–2023 were included in the study sample (94 cases in total). This retrospective cohort study assessed survival time (in days) from accident to death. Kaplan–Meier curves, stratified by key categorical variables, were used to analyze survival probabilities over time. Univariate Cox regression was used to assess each variable’s effect on survival. The association between exposure and mortality was analyzed using hazard ratios (HRs) and 95% confidence intervals (CIs). Results: Head injuries were associated with poorer outcomes. Chest injuries reduced median survival to 1.68 h compared to 5.85 days in cases that had not sustained chest injuries. Abdominal injuries also shortened survival (1.632 h vs. 1.896 h), as did multiple-site injuries (1.584 h vs. 0.2 days for single/double-site). Positive toxicology for psychoactive substances lowered survival to 1.32 h compared to 1.752 h in cases with negative toxicological findings. Multiple-site injuries and head, chest and abdominal injuries appeared to significantly affect the survival of victims. Positive toxicological examination results for psychoactive substances also appeared to heavily impact survival.

Forensic Sciences doi: 10.3390/forensicsci5040067

Authors: Alessia Leggio Giulia Iacobellis Cecilia Salzillo Liliana Innamorato

Background/Objectives: Establishing the identity of unknown individuals has always been one of the primary objectives of anthropologists and forensic pathologists in judicial contexts. Particularly when human remains are found in advanced stages of decomposition, carbonization, or fragmentation conditions that may compromise the efficacy of techniques such as DNA analysis or dental comparison innovative methodologies, including craniofacial superimposition, are employed, often supplemented by further examinations. This study presents the discovery of an individual in an advanced state of decomposition, transitioning from the colliquative to the semi-skeletal phase, demonstrating how degenerative processes can alter soft tissues to the extent of hindering genetic investigations. Methods: The multidisciplinary investigation conducted to resolve the case is described in two phases: the first, of an anthropological and medico-legal nature, aimed at reconstructing the biological profile (sex, age, stature, ancestry); the second, anthropological in focus, directed toward identification through craniofacial superimposition, applying two established methods from the literature the linear method and the computer-assisted comparison approach. Results: The results obtained from both investigative phases proved decisive, providing a significant and anticipated resolution for the authorities involved. Conclusions: This judicial case ultimately reaffirms the critical importance of multidisciplinary collaboration in forensic investigations.

Forensic Sciences doi: 10.3390/forensicsci5040066

Authors: Suthat Duangchit Naphatchaya Imkrajang Worrawit Boonthai Nareelak Tangsrisakda Sararat Innoi Sitthichai Iamsaard Chanasorn Poodendaen

Background/Objectives: Determination of the biological profile, particularly sex and stature, constitutes an essential component for individual identification in forensic and archaeological anthropology; however, validation of anthropometric equations remains inadequately implemented in contemporary research. This study addresses two limitations: the isolated development of sex and stature estimation methods and the lack of rigorous validation using independent samples. Methods: In this research, we analyzed 400 well-preserved scapulae from a Northeastern Thai population divided into training (n = 300: 150 male scapulae, 150 female scapulae) and validation (n = 100: 50 male scapulae, 50 female scapulae) groups. Eight standardized measurements were used for both sex and stature estimation. Results: All measurements demonstrated significant sexual dimorphism, with males exhibiting larger dimensions. For sex estimation, a multivariate model incorporating the maximum scapular height, maximum length of the spine, and scapula weight achieved 96.3% accuracy in the training samples and maintained 95.0% accuracy in independent validation. For stature estimation, a three-variable equation combining scapula weight, longitudinal scapular length, and maximum scapular breadth was strongly correlated (R = 0.769, SEE = 5.32 cm) with consistent performance across validation samples. Conclusions: Validation testing confirmed the high accuracy, reliability, and stability of both equations when applied to independent samples, with no significant differences in performance metrics between training and validation groups. These validated equations provide reliable standards for forensic practitioners analyzing scapular remains in practical applications within the northeastern Thai population.

Forensic Sciences doi: 10.3390/forensicsci5040065

Authors: Vittoria Medici Nicola Paone Giuseppe Pandarese Giuseppe Riccio Vito Alessandro Spinelli Gaetano Rizza Massimiliano Olivieri Milena Martarelli

Background: The analysis of gunshot residue (GSR) is crucial for gaining information on how a crime occurred. This study presents an innovative proof of concept for measuring shooting distances by performing Flash-Pulse active Thermography (FPT). Compared to conventional chemical methods, FPT offers a significant advantage by digitalizing the residue pattern in a non-destructive manner. Methods: Thermal images of cotton canvases, both white and colored, were analyzed to quantify the distribution of gunshot residues after shooting from several distances, specifically focusing on smoke and semi-burnt powders. The proposed approach uses contrast and radial intensity profiles to extract exponential coefficients, which are dependent on the shooting distance. Results: Employing a sigmoid model to fit the coefficients over distance and to derive a characteristic feature used as a classification metric, firing distances can be classified into short, medium, and long range and can be predicted with an uncertainty of less than 5 cm for distances between 18 and 38 cm under the tested conditions. Considerations regarding the influence of different weapons and ammunition are reported, suggesting the potential for a general approach. Conclusions: The methodology has been validated on several samples, demonstrating its feasibility for specific forensic applications. Its most robust use is as a weapon- and ammunition-specific calibration tool, supporting case-specific distance estimation analysis.

Forensic Sciences doi: 10.3390/forensicsci5040064

Authors: Karen M. Caulfield Nicola S. Gray Andrew Edwards Robert J. Snowden

Background: The DASH risk assessment scheme is used across the UK to identify and manage instances of domestic abuse. Recent studies have questioned whether the scheme can identify offenders who go on to commit further acts of domestic abuse, in particular serious violence, and therefore whether it is fit for purpose. Methods: We therefore tested the ability of the DASH to predict future instances of deadly or persistent domestic abuse. From a database of ≈25,000 incidents, we compared DASH assessments which preceded an incident of “deadly violence” or was the first in a series of “persistent abuse”. These groups were compared to a control group where there was no further incident of domestic abuse. Results: The proportion of “high-risk” stratifications was approximately 5 times higher in the deadly violence group compared to the control group. Prediction accuracy assessed via signal detection theory showed the DASH was a moderate predictor of deadly violence (AUC = 0.67). The DASH also showed predictive accuracy in identifying persistent offenders (AUC = 0.62). While these results are encouraging and are similar in efficacy to other risk assessment schemes used in the prediction of domestic violence, the results identified that many individual items of the DASH were not predictive. The inclusion of non-predictive items within the DASH adds “noise” and error into the risk evaluation. The development of a shortened version of the DASH, removing these ineffectual items, was shown to have even higher predictive value for deadly violence (AUC = 0.80). Conclusions: We stress, however, that the role of risk assessment is not to predict violence per se, but to prevent violence via the accurate identification of dangerous perpetrators and via effective intervention and safeguarding of victims. Despite this, research such as this is imperative to evaluate if the risk assessment schemes selected by practitioners and police are fit for purpose.

Forensic Sciences doi: 10.3390/forensicsci5040063

Authors: Allison Uvelli Erica Pugliese Fabio Ferretti

Background/Objectives: Chronic pain in women arises from a complex interplay of biological, psychological, and social factors. Despite its impact, validated screens for these bio-psycho-social risk factors are lacking. This study aimed to develop and validate a new screening and assessment tool to prevent chronic pain onset and detect hidden experiences of violence or trauma in women, also supporting individualized treatment. Methods: Conducted from December 2023 to June 2024 as part of a larger project, the original instrument comprised 36 bio-psycho-social risk factors. Rasch analysis was used for validation, assessing Infit, Outfit, DIF, ROC curves, and reliability indexes. Results: The sample included 239 women (100 victims), with 103 experiencing chronic pain, and 136 pain-free. Seven items were excluded due to poor fit, resulting in a 29-item version that met Rasch model criteria. Conclusions: The validated 29-item checklist, available in Italian, supports the bio-psycho-social model by identifying risk factors for chronic pain onset and detecting violence-related variables in diagnosed individuals. It offers clinicians and operators a practical tool to guide prevention and tailor personalized treatments. In addition, the RFCP-CK holds forensic relevance by offering measurable indicators that can support medico-legal evaluations, especially in cases where chronic pain is claimed to be a consequence of violence.

Forensic Sciences doi: 10.3390/forensicsci5040062

Authors: Sangita Baruah R. Suresh Rajesh Babu Govindarajulu Chandan Jyoti Kumar Bibhakar Chanda Lakshya Dugar Manob Jyoti Saikia

Background/Objectives: Toolmark analysis on cartridge cases offers critical insights in forensic ballistics, as the impressions left on cartridge cases by firearm components—such as the firing pin, breech face, and annulus—carry distinctive patterns and act as unique identifiers that can be used for firearm linkage. This study aims to develop a systematic and interpretable feature extraction pipeline for these regions to support future automation and comparison studies in forensic cartridge case analysis. Methods: A dataset of 20 high-resolution cartridge case images was prepared, and each region of interest (firing pin impression, breech face, and annulus) was manually annotated using the LabelMe tool. ImageJ and Python-based scripts were employed for feature extraction, capturing geometric descriptors (area, perimeter, circularity, and eccentricity) and texture-based features (Local Binary Patterns and Haralick statistics). In total, 61 quantitative features were derived from the annotated regions. Similarity between cartridge cases was evaluated using Euclidean distance metrics after normalization. Results: The extracted and calibrated region-wise geometric and texture features demonstrated distinct variation patterns across firing pin, breech face, and annulus regions. Pairwise similarity analysis revealed measurable intra-class differences, indicating the discriminative potential of the extracted features even within cartridges likely fired from the same firearm. Conclusions: This study provides a foundational, region-wise quantitative framework for analysing cartridge case impressions. The extracted dataset and similarity outcomes establish a baseline for subsequent research on firearm identification and model-based classification in forensic ballistics.

Forensic Sciences doi: 10.3390/forensicsci5040061

Authors: Beatrice Benedetti Caterina Pesaresi Stefania Molent Luigi Carbone Fabio De-Giorgio

Background: Cancer-related cachexia remains a significant cause of death, particularly for undiagnosed or untreated malignancies. Lymphomas, especially in uncommon locations, may go unrecognized until their advanced stages. Methods: We report the case of a 34-year-old woman who died from cancer-related cachexia due to undiagnosed, untreated cervical non-Hodgkin lymphoma. Four months postpartum, she reported having excluded malignancy through medical investigations, which were later confirmed to have never been performed. The Judicial Authority ordered an autopsy to determine the cause of death. A narrative literature review was conducted via PubMed using the terms “Non-Hodgkin Lymphoma” and “Autopsy,” limited to English-language human studies published between January 2000 and February 2025. Results: At autopsy, marked fat depletion and a 1350 g cervical mass were found, with significant anatomical distortion and airway narrowing due to epiglottic edema. Microscopic examination identified a diffuse large B-cell lymphoma of germinal center origin. A literature review on lymphoma-related autopsy findings identified common diagnostic challenges, including nonspecific symptoms, rapid clinical deterioration, the rarity of certain subtypes, and a lack of medical compliance. Conclusions: Early recognition and proper investigation of lymphoproliferative disorders are crucial to prevent fatal outcomes. Postmortem findings can offer valuable insights into missed diagnoses and inform strategies to reduce diagnostic delay.

Forensic Sciences doi: 10.3390/forensicsci5040060

Authors: Joe Adserias-Garriga Heli Maijanen Sara C. Zapico

Background: Sex estimation is a basic step of human identification in both legal cases and archeological contexts. The highest accuracy for sex estimation is achieved when a complete skeleton is available, though there are situations, such as cremated, dismembered, and otherwise taphonomically altered skeletal remains, where a complete skeleton is not available. The aim of the present preliminary study was to evaluate the usefulness of four non-metric skull traits that are considered taphonomically resilient for sex estimation and their potential application in forensic cases. Methods: Non-metric skull traits of 100 skulls from the Bass Donated Skeletal Collection were analyzed. These traits included foramen magnum shape, zygomatic arch extension with respect to the external auditory canal, sigmoid notch, and gonial angle muscle attachment. A discriminant function analysis model was used to develop specific formulae for sex estimation. Results: The foramen magnum and sigmoid notch showed no significant differences between males and females. The zygomatic arch extension (ZAE) and gonial angle morphology (GO) showed strong, significant differences between the sexes. However, gonial angle morphology has shown to be affected by edentulism. Based on the ZAE, the function obtained by the discriminant function analysis was sex = 2.469*ZAE − 1.247, with a result of zero pointing to males and result of one pointing to females, which correctly classified 79.8% of the original cases. Conclusions: This study highlights the value of four different skull traits and their potential use in forensic cases. Of all the evaluated traits, zygomatic arch extension was the best indicator for sex estimation. This anatomical region corresponds to a highly resistant skeletal structure.

Forensic Sciences doi: 10.3390/forensicsci5040059

Authors: Patrycja Myszak Laura Leclair Olivier Khayat Joshua Levy Joseph Abou Jaoude Mathieu Dufour Stéphanie Borduas Pagé Alexandre Hudon

Background/Objectives: The Commission d’examen des troubles mentaux (CETM), under Quebec’s Tribunal Administratif du Québec, reviews individuals found not criminally responsible on account of mental disorder (NCRMD). These hearings seek to balance public safety with reintegration, guided largely by treatment team recommendations. Despite the CETM’s central role in forensic psychiatry, limited empirical data exist on how its decisions align with clinical advice and which dynamic risk factors influence outcomes. This study aimed to (1) profile the CETM’s 2023 caseload, (2) evaluate concordance between CETM dispositions and treatment team recommendations, and (3) examine clinical, social, and legal factors associated with decision-making. Methods: We conducted a retrospective review of 1721 judgments issued by the CETM in 2023, retrieved from the publicly accessible Société Québécoise d’information juridique (SOQUIJ) database. Eligible cases included annual NCRMD review hearings, excluding trial fitness assessments and repeated hearings within the same year. A structured coding grid documented sociodemographic, administrative, legal, and clinical information, with emphasis on dynamic risk factors such as treatment adherence, substance use, and recent aggression. Descriptive analyses summarized population characteristics and concordance between clinical recommendations and CETM decisions. Results: The cohort was predominantly male (85%) with a mean age of 41 years. Psychotic disorders were the most frequent primary diagnoses (76%), frequently accompanied by substance use and antisocial traits. Most patients (79.6%) had prior psychiatric hospitalizations, while 25.5% had prior incarcerations. Nearly half displayed recent aggression or non-compliance. Treatment teams most often recommended conditional discharge (55%), followed by detention with conditions (21%) and unconditional release (19%). CETM decisions aligned with recommendations in 83.6% of cases; when divergent, rulings were more restrictive (8.6%) than permissive (4.6%). Conclusions: This study provides the first large-scale profile of Quebec’s CETM. High concordance with clinical teams was observed, but restrictive decisions were more frequent in cases of disagreement. The findings underscore the importance of incorporating standardized risk assessment tools to enhance transparency, consistency, and balance in forensic decision-making.

Forensic Sciences doi: 10.3390/forensicsci5040058

Authors: Naoto Tani Kazunori Miyamoto Takaki Ishikawa

Background: The present exploratory study aimed to investigate the relationship between arginine vasopressin (AVP) secretion and cerebral edema and evaluate its usefulness as an index for assessing severity of cerebral edema. Methods: Forensic autopsy was performed within 96 h after death in 143 cases, and samples of right heart blood and cerebrospinal fluid (CSF) were collected. Serum AVP levels and CSF were then measured and relationships with brain weight (BW) and normalized BW were investigated. Results: No relationships between serum AVP levels or CSF and age, sex, survival, or postmortem period were identified. A comparison in regard to cause of death revealed lower CSF AVP concentrations in cases of blunt head injury and cerebrovascular disease. In addition, a correlation was observed between serum AVP levels and non-normalized or normalized BW with blunt head injury or asphyxia. Receiver operating characteristic analysis revealed that neither serum nor CSF AVP concentrations yielded cutoff values readily applicable to the diagnosis of cerebral edema. Conclusions: The present findings suggest that postmortem AVP concentrations remain relatively stable and may be involved in the formation of cerebral edema. The findings also highlighted issues such as the influence of confounding factors associated with individual pathologies and the verification of AVP stability in postmortem samples. Thus, the usefulness of AVP as an indicator of cerebral edema in forensic evaluation remains unclear.

Forensic Sciences doi: 10.3390/forensicsci5040057

Authors: Stephanie J. Cole Heather M. Garvin

Background/Objectives: Incomplete cut marks produced during dismemberment are often interpreted as indicative of saw class characteristics. However, empirical validation of these associations remains limited, with prior studies examining six or fewer saws. Considering the wide variety of saws available, it is critical to assess the reliability of reported relationships between kerf features and saw classification using a larger sample, particularly in light of the serious legal consequences of erroneous conclusions. This study examines the statistical relationships between five incomplete cut traits—kerf profile shape (KPS), kerf length shape (KLS), floor dip (FD), kerf flare (KF), and floor striae (FS)—and saw class characteristics, including tooth set, tooth shape, teeth-per-inch, power, handle orientation, and cut direction. Methods: Kerf features were scored on a sample of 472 incomplete cuts made with 34 power and hand saws. Results: In reciprocating saws, W-shaped KPS was exclusively associated with crosscut, alternating saws (100%; p < 0.001), with hourglass-shaped KLS also primarily made by alternating sets (95.6%). Necked KLS was linked to wavy sets (76.8%; p < 0.001). FD, though rare, could be correctly assigned to teeth-per-inch groups (86.4%), and was also predominantly associated with alternating saws (90.9%; p < 0.001). Undulating FS were indicative of alternating saws with less than 20 teeth-per-inch (100%, p < 0.001). In contrast, KF showed no strong relationship with saw class characteristics, including handle side. Conclusions: The results of this large-scale analysis support most reported relationships in the saw mark literature but challenge assumptions that KF reliably indicates handle orientation or cut direction, suggesting instead that its location may reflect sawyer technique.

Forensic Sciences doi: 10.3390/forensicsci5040056

Authors: Wazha Mmereki Rodrigo S. Jamisola Zoe C. Jewell Tinao Petso Oduetse Matsebe Sky K. Alibhai

Background/Objectives: This study investigated the use of artificial intelligence (AI) to identify and match barefoot prints belonging to the same individual on soft and sandy soil substrates. Recognizing footprints on soil is challenging due to low contrast and variability in impressions. Methods: We introduce Deep Learning Footprint Identification Technology (DeepFIT), based on a modified You Only Look Once (YOLOv11s) algorithm, using three methods, namely, Bounding Box (BBox), 16 anatomical landmarks, and automatically segmented outlines (Auto-Seg). An Extra Small Detection Head (XSDH) was added to improve feature extraction at smaller scales and enhance generalization through multi-scale supervision, reducing overfitting to specific spatial patterns. Results: Forty adults (20 males, 20 females) participated, with 600 images per individual. As the number of individuals in model training increased, the BBox model’s accuracy declined, resulting in misclassification on the test set. The average performance accuracy across both substrates was 77% for BBox, 90% for segmented outlines, and 96% for anatomical landmarks. Conclusions: The landmark method was the most reliable for identifying and matching barefoot prints on both soft and sandy soils. This approach can assist forensic practitioners in linking suspects to crime scenes and reconstructing events from footprint evidence, providing a valuable tool for forensic investigations.

Forensic Sciences doi: 10.3390/forensicsci5040055

Authors: Francesco Orsini Stefania De Simone Roberta Bibbò Giovanni Pollice Luigi Cipolloni Stefano Ferracuti

Background/Objectives: Overkill is generally defined as violence demonstrably exceeding the force required to cause death, often involving a disproportionate number of inflicted injuries. Typically linked to specific mental states, this phenomenon can serve diverse perpetrator purposes, from expressing sadistic impulses to deliberately contaminating forensic evidence. This study re-examines the historical case of Mary Jane Kelly, the canonical final victim of Jack the Ripper, to offer a historical perspective on extreme violence and its influence on modern forensic frameworks. Methods: We employed a retrospective analysis, combining a systematic literature review with application of the Homicide Injury Scale to documented autopsy findings from 1888. Results: The autopsy revealed systematic post-mortem mutilation following a fatal neck laceration. Application of the Homicide Injury Scale confirmed violence exceeding lethal force. Conclusions: This case represents a foundational example of organized overkill behavior, demonstrating the value of applying modern forensic frameworks to historical cases to inform comprehensive overkill analysis.

Forensic Sciences doi: 10.3390/forensicsci5040054

Authors: K. Godde Samantha M. Hens Gwendolyn Fuentes

Background/Objectives: The identification of a decedent through skeletal analysis is dependent on accurate estimation of demographic characteristics, including biological sex. The most well-known sex estimation technique using the pubic bone is the Phenice method. In 2012, it was revised by Klales and colleagues and a logistic regression equation to predict sex was applied. Later, a program that estimates sex from Klales’ scoring with a random forest model, MorphoPASSE, was developed by Klales. Methods: Here we compare the accuracy of the original and revised methods, along with MorphoPASSE, using a contemporary sample of Northern Italians with documented sex. We further test the assertions by Phenice that his method is easy to employ for new observers and that ambiguity can be applied when characteristics do not morphologically fit into the categories of the method. Accuracy, error, bias, sensitivity, and specificity were calculated for each approach, along with McNemar’s tests for paired data, which compared documented sex and estimated sex. A linear weighted Cohen’s Kappa measured the differences in scoring between a new observer and an experienced observer. Results: Phenice’s method achieved higher accuracy (97%) than the Klales method and MorphoPASSE (86% each), as well as higher sensitivity and specificity, and lower error and bias. All McNemar’s tests conducted were not significant. The new observer demonstrated a similar accuracy (93%) to the experienced observer (97%). Furthermore, comparisons of Phenice’s scoring with ambiguity indicate its superior performance for capturing variation over the Klales method and MorphoPASSE. Conclusions: Phenice’s method is recommended in forensic anthropology and bioarchaeological contexts, particularly in Milan.

Forensic Sciences doi: 10.3390/forensicsci5040053

Authors: Ido Hefetz

Background: Artificial intelligence is transforming forensic fingerprint analysis by introducing probabilistic demographic inference alongside traditional pattern matching. This study explores how AI integration reshapes the role of forensic experts from interpreters of physical traces to epistemic corridors who validate algorithmic outputs and translate them into legally admissible evidence. Methods: A conceptual proof-of-concept exercise compares traditional AFIS-based workflows with AI-enhanced predictive models in a simulated burglary scenario involving partial latent fingermarks. The hypothetical design, which does not rely on empirical validation, illustrates the methodological contrasts between physical and algorithmic inference. Results: The comparison demonstrates how AI-based demographic classification can generate investigative leads when conventional matching fails. It also highlights the evolving responsibilities of forensic experts, who must acquire competencies in statistical validation, bias detection, and explainability while preserving traditional pattern-recognition expertise. Conclusions: AI should augment rather than replace expert judgment. Forensic practitioners must act as critical mediators between computational inference and courtroom testimony, ensuring that algorithmic evidence meets legal standards of transparency, contestability, and scientific rigor. The paper concludes with recommendations for validation protocols, cross-laboratory benchmarking, and structured training curricula to prepare experts for this transformed epistemic landscape.

Forensic Sciences doi: 10.3390/forensicsci5040052

Authors: Vito Maria Goffredo Michela Ferrara Mariagrazia Calvano Natascha Pascale Aldo Di Fazio Giuseppe Bertozzi

Background/Objectives: Cannabis is the most widely used illicit substance worldwide, particularly among young adults, with growing acceptance following medical and recreational legalization. Although generally perceived as a drug with low acute toxicity, an expanding body of evidence indicates that cannabinoids can exert relevant cardiovascular effects, including arrhythmias, myocardial ischemia, and sudden cardiac death (SCD). These mechanisms are mediated through complex, dose-dependent interactions among CB1 and CB2 receptors, autonomic imbalance, and endothelial dysfunction. Nevertheless, cannabis-related fatalities remain underestimated in both clinical and forensic settings. Case presentation: Three cases of sudden unexpected death in previously healthy men aged 28, 37, and 37 years are described. All were found deceased at home under non-suspicious circumstances. Forensic autopsies ruled out trauma, coronary atherosclerosis, congenital malformations, or cardiomyopathy. Histological analyses consistently revealed polymorphic myocardial alterations, including interstitial edema, fiber disruption, and focal myocytolysis, without inflammatory infiltrates or necrosis. Toxicological examinations demonstrated the presence of Δ9-tetrahydrocannabinol (THC) and metabolites in peripheral blood and urine, while alcohol and other illicit drugs tested negative. In each case, the cause of death was attributed to arrhythmic sudden cardiac death in temporal association with cannabis use. Conclusions: This case series, integrated with a narrative review of current literature, supports the hypothesis that cannabis consumption can contribute to fatal arrhythmias even in young adults without conventional cardiovascular risk factors. The convergence of autopsy, histopathological, and toxicological findings suggests a potential causal link between THC exposure and sudden unexpected death. These results highlight the importance of systematic postmortem investigations in suspected drug-related fatalities and underscore the need for greater awareness among clinicians, forensic pathologists, and policymakers regarding the underestimated cardiovascular toxicity of cannabis.

Forensic Sciences doi: 10.3390/forensicsci5040051

Authors: Sarajane Smith-Escudero Jesse R. Goliath Anna J. Osterholtz D. Shane Miller

Missing persons is an understudied area, especially within forensic anthropology when not associated with cases of mass violence or mass disaster. Affecting more than half a million persons in the United States yearly, missingness has proven to be a significant issue across the country—and globally—with no explicit preventative measures or solutions having been identified. Here we contribute to conceptualizations regarding the incidence of going missing, or missingness, utilizing frameworks of structural violence, necropolitics, and intersectionality. The concept of missingness is then applied to a case study of missing persons from Mississippi, documented in the Mississippi Repository for Missing and Unidentified Persons. Missingness was found to affect demographic groups differently, with marginalized racial and ethnic groups not only disproportionately exposed to the risk of missingness but also suffering from the worst case outcomes after having gone missing. We argue that the socioeconomic, political, and sociocultural limitations structural violence imposes upon marginalized groups may result in missingness. By rigorously theorizing missingness, we hope to draw attention to and improve an understanding of the silent epidemic of missingness, eventually resulting in the identification of specific circumstances correlated to missingness and ultimately improving prevention and recovery efforts.

Forensic Sciences doi: 10.3390/forensicsci5040050

Authors: Raffaella Minella Giada Sciâdi Steiger Aldo Di Fazio Francesco Introna Enrica Macorano

Background/Objectives: Age estimation is of fundamental importance in forensic investigations. When traditional methods based on gross bone morphology or morphometric analysis cannot be applied, forensic experts must rely on multidisciplinary approaches. Histomorphometry has consistently proven to be reliable in cases of highly fragmented or incomplete skeletal remains, particularly in older individuals. Building on the foundational study of Amprino and Bairati, this study evaluated the correlations between bone microstructural features in femoral cross-sections and the age and sex of individuals. Methods: The sample comprised 95 femoral mid-diaphyseal thin sections obtained from autopsy specimens housed at the Institute of Legal Medicine, University of Bari (Italy), representing both male and female individuals aged 18 to 92 years. The numbers and densities of primary, intact secondary, and fragmentary secondary osteons, together with osteon circularity and the mean osteonal area, were measured to investigate age-related variation. Statistical analyses included t-tests, Mann–Whitney tests, Spearman’s rank correlation, and General Linear Models (GLMs). Results: No significant differences in histomorphometric variables were observed between males and females. However, the number of intact secondary osteons and osteon population density increased with age, while the mean osteonal area and osteon circularity decreased with age. Although some variables displayed significant correlations with age, residual analysis indicated a lack of heterogeneity in variance, which limited the development of a robust predictive model. Conclusions: The findings highlight both the potential and the limitations of histomorphometry in forensic age estimation. While certain microstructural variables correlate with age, inter-individual variability reduces predictive accuracy. Further research is needed to refine models that account for biological and biomechanical variability, particularly in older adults.

Forensic Sciences doi: 10.3390/forensicsci5040049

Authors: Mackenzie S. Todd Julieta G. García-Donas

Background/Objectives: Estimating the sex from unknown individuals is a critical step when constructing their biological profile. The distal humerus is a useful sex discriminator as shown through metric, morphoscopic, and geometric morphometric approaches. A recently developed web application using geometric morphometric techniques has provided an accessible tool for estimating sex from the shape of the olecranon fossa. The aims of this study were to examine the accuracy of the Ammer–Coelho web application on Scottish individuals, as well as test its repeatability and reproducibility among seven different observers. Methods: The right humerus was obtained from 52 Scottish individuals, and the Ammer–Coelho web application was used to estimate sex. Total accuracy rates and sex-specific rates were calculated, and an analysis of Cohen’s and Fleiss’ kappa was performed. Results: The results demonstrate an overall accuracy of 69.23% with a sex bias of −5.33%, with 55.56% of the sample being accurately estimated with probabilities equal to or higher than 0.95. Substantial agreement was reported for intra-observer error, and an overall low agreement was reported for inter-observer error Conclusions: This is the first study that evaluates the Ammer–Coelho web application. A tendency to perceive more triangular shapes (male appearance) rather than oval shapes (female appearance) resulted in a high level of observer errors, with only 6% of females correctly estimated across the seven observers. The low accuracy rates obtained could also indicate inter-population variation, as shown by other studies. Due to the results obtained, research considering different levels of observers’ experience and diverse population samples is needed to confirm our findings.

Forensic Sciences doi: 10.3390/forensicsci5040048

Authors: Romil Rawat Hitesh Rawat Mandakini Ingle Anjali Rawat Anand Rajavat Ashish Dibouliya

Background/Objectives—Traditional forensic investigations often analyze digital, physical, and criminological evidence separately, leading to fragmented timelines and reduced accuracy in reconstructing complex events. To address these gaps, this study proposes the Digital Stratigraphy Framework (DSF), inspired by archaeological stratigraphy, to integrate heterogeneous evidence into structured, temporally ordered layers. DSF aims to reduce asynchronous inconsistencies, minimize false associations, and enhance interpretability across digital, behavioral, geospatial, and excavation evidence. Methods—DSF employs Hierarchical Pattern Mining (HPM) to detect recurring behavioral patterns and Forensic Sequence Alignment (FSA) to synchronize evidence layers temporally and contextually. The framework was tested on the CSI-DS2025 dataset containing 25,000 multimodal, stratified records, including digital logs, geospatial data, criminological reports, and excavation notes. Evaluation used 10-fold cross-validation, Bayesian hyperparameter tuning, and structured train-validation-test splits. Metrics included accuracy, precision, recall, F1-score, and Stratigraphic Reconstruction Consistency (SRC), alongside ablation and runtime assessments. Results—DSF achieved 92.6% accuracy, 93.1% precision, 90.5% recall, 91.3% F1-score, and an SRC of 0.89, outperforming baseline models. False associations were reduced by 18%, confirming effective cross-layer alignment and computational efficiency. Conclusions—By applying stratigraphic principles to forensic analytics, DSF enables accurate, interpretable, and legally robust evidence reconstruction. The framework establishes a scalable foundation for real-time investigative applications and multi-modal evidence integration, offering significant improvements over traditional fragmented approaches.

Forensic Sciences doi: 10.3390/forensicsci5040047

Authors: Dalila Tripi Miriam Ottaviani Federica Spadazzi Maria Vittoria Zamponi Claudia Casella Giuseppe Bertozzi Raffaele La Russa Lina De Paola

Background: Adoption in Italy has evolved significantly over the years, becoming a critical legal, social, and ethical issue. Recent data indicates a decline in the number of adoptions, reflecting a broader European trend. Historically, adoption was primarily pursued by couples unable to have biological children due to infertility. Today, however, the adoption process in Italy has expanded to include single individuals, following recent legislative reforms that aim to accommodate diverse family structures. Methods: We have analyzed the relevant literature through a PubMed search and studied the current regulations considering recent Italian developments. This paper examines the current state of adoption in Italy, focusing on the increasing need for interdisciplinary collaboration between law, forensic medicine, ethics, and psychology. Results: The evolving adoption landscape requires a comprehensive approach that addresses legal issues, medical conditions, ethical considerations, and psychological support for both adoptive parents and children. The integration of these fields is essential for ensuring the well-being of the adopted child and the success of the adoption process. Conclusions: By exploring the dynamics of adoption, this article highlights the importance of a multidisciplinary framework in meeting the needs of children awaiting adoption and fostering the development of new family structures in Italy. In particular, forensic medicine plays a central role in evaluating parental suitability, detecting potential risk factors, and supporting judicial decisions through technical expertise and medico-legal assessments.

Forensic Sciences doi: 10.3390/forensicsci5030046

Authors: Katalin Fogarasi Gábor Simon Gábor Gyenes Péter Gergely Zoltán Patonai

Background/Objectives: Clinical documentation of injuries is essential for forensic evaluation, especially in criminal cases. However, vague descriptions and inconsistent terminology often reduce the forensic utility of these records. This study examines the terminological quality and forensic interpretability of clinical injury documentation with the aim of improving medico-legal assessments. Methods: A corpus of 1000 Hungarian medical diagnostic reports of injuries was analyzed using descriptive statistics and manual terminological review. Results: Significant gaps in morphological detail and frequent terminological inconsistencies were found. Terms describing incised and chop wounds were often used interchangeably or inaccurately, impairing forensic interpretation of injury mechanisms, instruments, severity, and anatomical location. These variations also challenge bodily harm classification and injury event reconstruction. Conclusions: The findings highlight the need for harmonizing clinical and forensic terminology. Improved consistency in injury documentation will enhance its clarity, reliability, and forensic value, facilitating better interdisciplinary collaboration and legal outcomes.

Forensic Sciences doi: 10.3390/forensicsci5030045

Authors: Kelly R. Kamnikar Nicollette S. Appel Hannah N. Cantrell Heather J. H. Edgar

Background/Objectives: This paper presents cranial MMS data for an American Indian (AI) sample from New Mexico. We compare these data to other population reference samples to understand biological distance and classification patterns. Methods: Cranial MMS data was collected from CT scans of AI individuals (n = 839) in the New Mexico Decedent Image Database. We used 12 traits following a published protocol for CT data and excluded nasal bone contour. The AI sample is compared to other samples of African American or Black, Asian, Hispanic, and White individuals to assess biological distance and classification accuracy. Results: Biological distance analysis indicated similarity among the AI, Hispanic, and Black samples, but dissimilarity among the Asian and White samples to the AI sample. Classification accuracy was low for the overall model, with Hispanic and Black individuals frequently misclassifying as AI. Conclusions: As is true everywhere, ideas about identity are complex in New Mexico. AI and Hispanic individuals in NM self-ascribe to one or both social groups. We found that biological data cannot meaningfully differentiate between these social groups, as crania are morphologically similar when examining cranial MMS traits. These results, coupled with New Mexican identity and genetics, contributes to the difficulty in group attribution in forensic casework. Nevertheless, the reference data for AI presented here can provide more robust statistics that support the likelihood of AI and Hispanic affinity in population affinity analysis. We underscore the need for an understanding of regional population history and structure and reference samples while assessing population affinity in forensic casework.

Forensic Sciences doi: 10.3390/forensicsci5030044

Authors: Maria Puntarello Ginevra Malta Mauro Midiri Tommaso D’Anna Giuseppe Davide Albano Stefania Zerbo Antonina Argo

Background/Objectives: Suicide in prison is a significant medico-legal and public health concern, with rates several times higher than in the general population. Vulnerability is heightened by psychiatric disorders, substance use, and custodial stressors such as isolation, overcrowding, and restricted healthcare access. This study examines custodial suicides to identify diagnostic complexities, systemic shortcomings, and possible prevention strategies, including technological innovations. Methods: We conducted a retrospective forensic investigation of sixteen confirmed custodial suicides between 2022 and 2024. Each underwent a standardized protocol comprising crime scene inspection, complete autopsy, histopathology, toxicology, and review of prison medical and psychiatric records. Data on suicide methods, psychiatric comorbidities, and substance use were analyzed. Results: Hanging was the predominant method (12/16), displaying classical forensic signs such as pale, oblique ligature marks and petechial hemorrhages. Four cases involved acute intoxication, often with non-prescribed drugs. Psychiatric disorders were identified in 14 cases, including major depressive disorder, bipolar disorder, and substance use disorder. Toxicological analyses revealed both prescribed and illicit substances, highlighting unauthorized exchanges within facilities. Autopsy findings consistently excluded homicide or natural causes, confirming the vitality of lesions and the mechanism of death. Conclusions: Custodial suicides are strongly associated with untreated or inadequately managed psychiatric conditions, compounded by restrictive prison environments. Comprehensive forensic autopsies are essential for accurate cause-of-death determination and institutional accountability. Preventive strategies should combine psychiatric care, architectural modifications to reduce ligature points, and ethical integration of AI-based surveillance for early detection of suicidal behavior. A multidisciplinary, rights-based approach is crucial to reduce suicide rates and safeguard the dignity and life of incarcerated individuals.

Forensic Sciences doi: 10.3390/forensicsci5030043

Authors: Kyra E. Stull Christopher A. Wolfe Briana T. New Louise K. Corron Kate Spradley

Background/Objectives: The incorporation of the human growth and development literature, an ontogenetic framework, a large virtual sample of individuals across the entire growth period, and a contemporary sample of adult individuals provides a unique opportunity to explore the cranial complex across the entire life cycle. This study (1) assesses cranial variation in postnatal ontogeny to determine the life history stage during which subadult crania can reach comparable levels of phenotypic expression to adult crania and (2) exposes when biological sex can be estimated using craniometric data from immature individuals with accuracy levels comparable to adults. Methods: Contemporary individuals between birth and 102 years of age from one virtual (Subadult Virtual Anthropology Database; SVAD) and one skeletal (Forensic Data Bank; FDB) collection were used in the analyses (n = 2152). Results: Discriminant analysis reveals a clear ontogenetic trajectory across the life history stages, with adolescents, SVAD adults, and FDB adults exhibiting similar cranial dimensions. The analysis also revealed a shift from the growth energetic period into the reproductive energetic period during adolescence. This transition is reflected in the divergence of male and female craniometrics in adolescence, which is also when sex estimation accuracy is comparable to SVAD and FDB adults. Conclusions: The current study argues that skeletal and/or dental maturity is not necessary to estimate sex using the cranium and urges the field to reconsider methodological divisions between subadults and adults.

Forensic Sciences doi: 10.3390/forensicsci5030042

Authors: Valentina Caparrelli Margherita Pallocci Roberta Tittarelli Carmelo Russo Laura Donato Francesca Ponzani Pierluigi Passalacqua Filippo Milano Michele Treglia

Suicide is a public health concern worldwide. The methods used worldwide can vary based on geographical context and the ease with which victims can find the means to act on suicidal thoughts. In this context, the recent emergence of an online market for toxic and narcotic substances has increased accessibility to these substances for recreational and suicidal purposes. The presented case concerns a fatal sodium nitrite dose purchased online. The body exhibited the characteristic blue-gray staining of hypostasis, more evident on the neck and the upper third of the posterior surface of the chest, and toxicological analysis revealed methemoglobin levels exceeding 30%. Although there is no standardized analytical procedure, the determination of nitrite in postmortem samples is crucial in these cases. Additionally, nitrite is difficult to detect postmortem because it rapidly turns into sodium nitrate. In the case presented, capillary ion analysis (CIA) with direct UV detection was performed for the qualitative and quantitative determination of nitrite and nitrate in the cadaveric central blood samples. The analytical determination identified the presence of nitrite ions and sodium nitrate, in a concentration of 9515 µmol/L.

Forensic Sciences doi: 10.3390/forensicsci5030041

Authors: Elaine Y. Chu Amariah J. Treviño Marissa E. Vasquez

Background/Objectives: Methods of estimating the biological profile in forensic anthropology must meet criteria set forth by standards for admissibility in legal proceedings. Subadult biological profile methods have not been as extensively validated due to limited sources of subadult skeletal reference data. Methods: Data for the contemporary Japanese validation sample were collected from full-body, postmortem computed tomography (PMCT) scans (n = 118). Ten subadult age and stature estimation methods using long bone lengths were validated for accuracy, precision, and bias. The methods included both linear and nonlinear regression. Results: Nonlinear regression methods yielded high validation accuracy and precision for age (>90%; <2.5 cm) and stature (>95.89%; <17 cm) and performed better than linear regression methods. Most methods do not meet criteria set by the American Academy of Forensic Sciences (AAFS) Standards Board (ASB) or the American National Standards Institute (ANSI). Conclusions: As skeletal data become increasingly accessible, it is important to continue to validate currently available methods for estimating aspects of the subadult biological profile while also prioritizing the creation of new population-specific and generic methods applicable for forensic casework. Particular focus should lie on improving reference skeletal material variation, appropriate statistical modeling, and adherence to standards in forensic anthropology. Recommendations for choosing the most appropriate method, given a subadult forensic case, are provided.

Forensic Sciences doi: 10.3390/forensicsci5030040

Authors: Livio Pietro Tronconi Vittorio Bolcato Luca Bianco Prevot Alessandro Carrozzo Giulia Bambagiotti Giuseppe Basile

Informed consent is a fundamental component of both the right to health and self-determination. Its violation, which occurs when the acquisition process is absent or flawed, can also lead to legal proceedings for isolated harm, regardless of the simultaneous existence of those related to biological and psychological harm. Across Europe, this remains a legal and ethical cornerstone, increasingly important in the context of cross-border care and culturally diverse healthcare settings. Many countries have enacted specific laws to ensure patients receive adequate information, while courts continue to refine its interpretation—often highlighting the inadequacy of informed consent in daily clinical practice. In Italy, in particular, recent rulings underscore the need for clear, comprehensive, and properly documented consent formats for health treatments. This review synthesizes rulings, regulations, and the scientific literature to offer practical guidance for improving informed consent practices in Italy, valuing the role of forensic experts in the integration in healthcare settings of the different interests at stake. The aim is to support both legal compliance and the delivery of high-quality, patient-centred care. Courts and legislation in Italy demand clear and tailored informative disclosure for health treatments, with indications of the treatment and outcomes, to be in written form. Yet, foreseeable risks, more common complications, and alternative treatment options are often poorly communicated in practice—frequently forming the basis of claims for compensation. Integrating team-based consultations and multimedia tools can significantly enhance patient understanding and secure the informed consent process. Those key points are summarized in a brief table, ideally suggesting the minimal requirements for an informed consent form. Investing in standardizing informed consent protocols, and also fostering the sharing of best practices with forensic medicine experts, are crucial steps in ensuring shared decision-making in healthcare and trying to reduce legal disputes.

Forensic Sciences doi: 10.3390/forensicsci5030039

Authors: Salvatore Nigliaccio Davide Alessio Fontana Emanuele Di Vita Marco Piraino Pietro Messina Antonina Argo Stefania Zerbo Davide Albano Enzo Cumbo Giuseppe Alessandro Scardina

Digital dentistry is undergoing rapid evolution, with three-dimensional imaging technologies increasingly integrated into routine clinical workflows. Originally developed for accurate dental arch reconstruction, modern intraoral scanners have demonstrated expanding versatility in capturing intraoral mucosal as well as perioral cutaneous structures. Concurrently, photogrammetry has emerged as a powerful method for full-face digital reconstruction, particularly valuable in orthodontic and prosthodontic treatment planning. These advances offer promising applications in forensic sciences, where high-resolution, three-dimensional documentation of anatomical details such as palatal rugae, lip prints, and bite marks can provide objective and enduring records for legal and investigative purposes. This study explores the forensic potential of two digital acquisition techniques by presenting two cadaveric cases of animal bite injuries. In the first case, an intraoral scanner (Dexis 3600) was used in an unconventional extraoral application to directly scan skin lesions. In the second case, photogrammetry was employed using a digital single-lens reflex (DSLR) camera and Agisoft Metashape, with standardized lighting and metric scale references to generate accurate 3D models. Both methods produced analyzable digital reconstructions suitable for forensic archiving. The intraoral scanner yielded dimensionally accurate models, with strong agreement with manual measurements, though limited by difficulties in capturing complex surface morphology. Photogrammetry, meanwhile, allowed for broader contextual reconstruction with high texture fidelity, albeit requiring more extensive processing and scale calibration. A notable advantage common to both techniques is the avoidance of physical contact and impression materials, which can compress and distort soft tissues, an especially relevant concern when documenting transient evidence like bite marks. These results suggest that both technologies, despite their different origins and operational workflows, can contribute meaningfully to forensic documentation of bite-related injuries. While constrained by the exploratory nature and small sample size of this study, the findings support the viability of digitized, non-destructive evidence preservation. Future perspectives may include the integration of artificial intelligence to assist with morphological matching and the establishment of digital forensic databases for pattern comparison and expert review.

Forensic Sciences doi: 10.3390/forensicsci5030038

Authors: Paola Frati Stefania De Simone Giovanni Pollice Elena Giacani Luigi Cipolloni Francesco Orsini

Background: Sexual violence poses a significant challenge to European lawmakers, impacting the victim’s physical and psychological health. This study examines sexual violence legislation across EU member states, Switzerland, and the UK, analyzing similarities, differences, challenges, and potential solutions for effective policy development. The research was motivated by the adoption of EU Directive 2024/1385. Methods: This study analyzes sexual violence legislation across European countries in a comparative and qualitative way, highlighting differences, commonalities, and the potential for uniform regulation. The data were collected from the literature published between 2015 and 2024, focusing the EU member states, Switzerland, and the United Kingdom. Results: The examination of the norms governing sexual offenses in various European countries revealed significant differences in legislative frameworks, reflecting diverse cultural, ethical, and legal perspectives. Conclusions: Despite European countries sharing the goal of protecting victims and combating sexual violence, there are significant legislative disparities. Key recommendations include enhancing EU member state cooperation, implementing joint training programs, developing a specific EU directive, and creating coordinated prevention and education programs. While respecting national legal diversity, a unified approach is needed for effective prevention and prosecution of sexual violence across Europe.

Forensic Sciences doi: 10.3390/forensicsci5030037

Authors: Maria João Couto Áurea Madureira-Carvalho Inês Morais Caldas

Background: In forensic anthropology, estimating stature is an essential part of constructing the biological profile of unknown individuals. While long bones are typically used for this purpose, they are often missing or incomplete in forensic contexts. Methods: This study examined the relationship between cranial and mandibular measurements and estimated stature in a sample of 84 identified adult (aged over 18 years) Portuguese skeletons (43 females and 41 males) from two osteological collections. Stature was estimated using Mendonça’s regression model based on humeral length. Four cranial and mandibular measurements were obtained, and intra- and interobserver reliability was assessed. Results: All variables showed statistically significant correlations with stature, although only the mandibular measurement—the distance between the mental symphysis and the mental foramen (SMFM)—and sex contributed significantly to the final regression model. The model explained 51.3% of the variance in stature. Conclusions: These findings suggest that SMFM, a stable mandibular marker, may serve as a useful supplementary indicator for stature estimation in cases where long bones are not available. Further studies with larger and more diverse samples are needed to validate these findings and evaluate their applicability across different populations.

Forensic Sciences doi: 10.3390/forensicsci5030036

Authors: Serena De Cunto Rosa Maria Di Maggio Pier Matteo Barone

The disappearance of foreign minors in Italy is a long-standing and critically underexamined social phenomenon. Despite alarming figures, public and institutional attention remains episodic and media-driven, often limited to high-profile or criminal cases. This study offers a socio-forensic analysis of official data from 2014 to 2023, revealing significant inconsistencies in how these cases are reported, categorized, and followed up by Italian institutions. It highlights how unaccompanied and migrant minors are especially vulnerable within a fragmented and reactive system that lacks transparency and effective preventive measures. Rather than presenting new empirical data, the article reinterprets existing sources to expose systemic gaps, drawing comparisons with the more structured approaches adopted in countries such as the United States, United Kingdom, and Spain. These international examples show how multilingual communication, early warning systems (e.g., AMBER Alert), and public geolocation tools can offer timely, coordinated responses to disappearances—tools that remain largely absent or underused in Italy. The article further argues for the integration of forensic geospatial methods, such as locus operandi analysis, remote sensing, and forensic geoarchaeology, not as experimental techniques, but as practical tools that could strengthen Italy’s institutional capacity to respond. Ultimately, this study seeks to elevate the discussion surrounding missing foreign minors from a marginal social concern to a matter of forensic and public interest, and to encourage interdisciplinary reflection on how such disappearances are framed—and too often dismissed—within the national landscape.

Forensic Sciences doi: 10.3390/forensicsci5030035

Authors: Joe Adserias-Garriga Hannah Skropits Brailey Moeder

Background: Positive identification is at the forefront of tasks for forensic practitioners when a set of remains is discovered. Standard means of identification include fingerprints, dental, and DNA analyses; however, additional methods are utilized by forensic practitioners to identify remains when these primary methods of identification are not applicable. Comparative radiography has become a frequently employed approach for positive identification, specifically focused on individualizing characteristics evident in human skeletal variation. Regions that display wide ranges of morphological variation within the human skeleton include the cranium as well as the thorax. With regard to the cranium specifically, paranasal sinuses have been recognized as unique features and are valuable for identification purposes. Objectives: This paper explores the basic information of the anatomy and development, range of variation, and the importance of paranasal sinuses in forensic contexts. Results: This article discusses how practitioners can best use the morphological information contained in the paranasal sinuses and how to compare the antemortem and postmortem datasets involving different imaging modalities for positive identification purposes, in order to provide practical concepts that may assist in cases where paranasal sinuses may be used for forensic human identification. Conclusions: Understanding the development of paranasal sinuses, the imaging techniques applied for their visualization, as well as the principles of identification, is key to conducting proper antemortem vs. postmortem comparisons and effectively utilizing paranasal sinuses in forensic identification contexts.

Forensic Sciences doi: 10.3390/forensicsci5030034

Authors: Pier Matteo Barone Enrico Di Luise

This case report presents a multidisciplinary forensic investigation into a cold case involving a missing person in Italy, likely linked to a homicide that occurred in 2008. The investigation applied a standardized protocol integrating satellite imagery analysis, site reconnaissance, vegetation clearance, ground-penetrating radar (GPR), and cadaver dog (K9) deployment. A dedicated decision tree guided each phase, allowing for efficient allocation of resources and minimizing investigative delays. Although no human remains were recovered, the case demonstrates the practical utility and operational robustness of a structured, evidence-based model that supports decision-making even in the absence of positive findings. The approach highlights the relevance of “negative” results, which, when derived through scientifically validated procedures, offer substantial value by excluding burial scenarios with a high degree of reliability. This case is particularly significant in the Italian forensic context, where the adoption of standardized search protocols remains limited, especially in complex outdoor environments. The integration of geophysical, remote sensing, and canine methodologies—rooted in forensic geoarchaeology—provides a replicable framework that enhances both investigative effectiveness and the evidentiary admissibility of findings in court. The protocol illustrated in this study supports the consistent evaluation of large and morphologically complex areas, reduces the risk of interpretive error, and reinforces the transparency and scientific rigor expected in judicial settings. As such, it offers a model for improving forensic search strategies in both national and international contexts, particularly in long-standing or high-profile missing persons cases.

Forensic Sciences doi: 10.3390/forensicsci5030033

Authors: Francesco Orsini Giovanni Pollice Francesco Carpano Luigi Cipolloni Andrea Cioffi Camilla Cecannecchia Roberta Bibbò Stefania De Simone

Background/Objectives: Traumatic brain injury is one of the leading causes of death and disability. When traumatic brain injury is repeated over time, it can lead to the development of Chronic Traumatic Encephalopathy, a chronic neurodegenerative disease commonly observed in individuals who engage in contact sports or military personnel involved in activities with a high risk of repeated head trauma. At autopsy, the examination of the brain reveals regional atrophy, corresponding to high concentrations of glutamate receptors. Microscopically, the primary findings are the deposition of neurofibrillary tangles and neuropil threads. The aim of this study is to highlight the clinical and histopathological characteristics of Chronic Traumatic Encephalopathy, providing diagnostic support to forensic pathologists. Additionally, it seeks to aid in the differential diagnosis of similar conditions. Methods: A review of literature was conducted following the PRISMA criteria. Of 274 articles, 7 were selected. Results: According to these papers, most patients were male and exhibited neurological symptoms and neuropsychiatric impairments, and a proportion of them committed suicide or had aggressive behavior. Conclusions: Chronic Traumatic Encephalopathy remains largely underdiagnosed during life. The definitive diagnosis of Chronic Traumatic Encephalopathy is established post-mortem through the identification of pathognomonic tauopathy lesions. Early and accurate antemortem recognition, particularly in at-risk individuals, is highly valuable for its differentiation from other neurodegenerative conditions, thereby enabling appropriate clinical management and potential interventions.

Forensic Sciences doi: 10.3390/forensicsci5030032

Authors: Briana T. New Kyra E. Stull Louise K. Corron Christopher A. Wolfe

Although cranial growth has been extensively explored, forensic and biological anthropology lack a formal incorporation of how cranial growth processes impact the adult phenotype and downstream biological profile estimations. Objectives: This research uses an ontogenetic framework to identify when interlandmark distances (ILDs) stabilize during growth to reach adult levels of variation and to evaluate patterns of cranial sexual size dimorphism. Methods: Multivariate adaptive regression splines (MARS) were conducted on standardized cranial ILDs for 595 individuals from the Subadult Virtual Anthropology Database (SVAD) and the Forensic Data Bank (FDB) aged between birth and 25 years. Cross-Validated R-squared (CVRSq) values evaluated ILD variation explained by age while knot placements identified meaningful changes in ILD growth trajectories. Results: Results reveal the ages at which males and females reach craniometric maturity across splanchnocranium, neurocranium, basicranium and cross-regional ILDs. Changes in growth patterns observed here largely align with growth milestones of integrated soft tissue and skeletal structures as well as developmental milestones like puberty. Conclusions: Our findings highlight the variability in growth by sex and cranial region and move forensic anthropologists towards recognizing cranial growth as a mosaic, continuous process with overlap between subadults and adults rather than consistently approaching subadult and adult research separately.

Forensic Sciences doi: 10.3390/forensicsci5030031

Authors: Mario Giuseppe Chisari Massimiliano Esposito Salvatore Alloca Sabrina Franco Martina Francaviglia Gianpietro Volonnino Raffaella Rinaldi Nicola Di Fazio Lucio Di Mauro

Background: Anabolic–androgenic steroids (AASs) are commonly used for performance enhancement but have been linked to significant neurobiological consequences. This review explores the impact of AASs on neurochemical pathways, cognitive function, and psychiatric disorders, highlighting their potential neurotoxicity. Methods: A narrative review of current literature was conducted to examine AASs-induced alterations in neurotransmitter systems, structural and functional brain changes, and associated psychiatric conditions. The interplay between AASs use and other substances was also considered. Results: Chronic AASs exposure affects serotonin and dopamine systems, contributing to mood disorders, aggression, and cognitive deficits. Structural and functional changes in the prefrontal cortex and limbic regions suggest long-term neurotoxicity. AASs use is associated with increased risks of depression, anxiety, and psychosis, potentially driven by hormonal dysregulation and neuroinflammation. Co-occurring substance use exacerbates neurocognitive impairments and behavioral disturbances. Discussion: While evidence supports the link between AASs use and neurotoxicity, gaps remain in understanding the precise mechanisms and long-term effects. Identifying biomarkers of brain damage and developing targeted interventions are crucial for mitigating risks. Increased awareness among medical professionals and policymakers is essential to address AASs-related neuropsychiatric consequences. Conclusions: AASs abuse poses significant risks to brain health, necessitating further research and prevention efforts. Evidence-based strategies are needed to educate the public, enhance early detection, and develop effective interventions to reduce the neuropsychiatric burden of AASs use.

Forensic Sciences doi: 10.3390/forensicsci5030030

Authors: Francesco Sessa Mario Chisari Massimiliano Esposito Elisa Guardo Lucio Di Mauro Monica Salerno Cristoforo Pomara

Background/Objectives: Artificial intelligence (AI) is beginning to be applied in wound ballistics, showing preliminary potential to improve the accuracy and objectivity of forensic analyses. This review explores the current state of AI applications in forensic firearm wound analysis, emphasizing its potential to address challenges such as subjective interpretations and data heterogeneity. Methods: A systematic review adhering to PRISMA guidelines was conducted using databases such as Scopus and Web of Science. Keywords focused on AI and GSW classification identified 502 studies, narrowed down to 4 relevant articles after rigorous screening based on inclusion and exclusion criteria. Results: These studies examined the role of deep learning (DL) models in classifying GSWs by type, shooting distance, and entry or exit characteristics. The key findings demonstrated that DL models like TinyResNet, ResNet152, and ConvNext Tiny achieved accuracy ranging from 87.99% to 98%. Models were effective in tasks such as classifying GSWs and estimating shooting distances. However, most studies were exploratory in nature, with small sample sizes and, in some cases, reliance on animal models, which limits generalizability to real-world forensic scenarios. Conclusions: Comparisons with other forensic AI applications revealed that large, diverse datasets significantly enhance model performance. Transparent and interpretable AI systems utilizing techniques are essential for judicial acceptance and ethical compliance. Despite the encouraging results, the field remains in an early stage of development. Limitations highlight the need for standardized protocols, cross-institutional collaboration, and the integration of multimodal data for robust forensic AI systems. Future research should focus on overcoming current data and validation constraints, ensuring the ethical use of human forensic data, and developing AI tools that are scientifically sound and legally defensible.

Forensic Sciences doi: 10.3390/forensicsci5030029

Authors: Petra Švábová Darina Falbová Zuzana Kozáková Mária Chovancová Lenka Vorobeľová Radoslav Beňuš

In forensic sciences, particularly in forensic anthropology and podiatry, assessing a person’s stature helps create a biological profile that allows for more accurate identification. Background/Objectives: When considering dynamic footprints as part of the gait pattern, certain parameters such as stride length, step length, gait width, and gait angle can be evaluated in relation to stature. The aim of this study was to assess footstep and stride length from the gait of dynamic footprints and determine if they correlate with stature and could be useful for biological profiling. Methods: Gait patterns from dynamic footprints and stature were determined in 114 females and 104 males aged 18 to 33 years. Results: All participants took the first step with their preferred foot, 56% with the right foot. Regarding step sequence, there were non-significant differences between the 4th and 5th footsteps in both sexes. Sex differences were significant in four of seven footsteps. Only a few steps significantly correlated in sequence with stature, and even these had low correlation coefficients (r = 0.295). In females, positive values of mean differences between actual and estimated stature predictions indicate that the equations tend to overestimate, whereas in a mixed sex group, most negative values of mean differences indicate underestimation. Conclusions: Given the weak correlations observed, footstep and stride length should not be considered reliable indicators for forensic stature estimation. These parameters are more suitable for biomechanical and anthropological research, while forensic applications should be considered supplementary and interpreted with caution.

Forensic Sciences doi: 10.3390/forensicsci5030028

Authors: Joel E. R. Moss Kathryn Hamory Robert Moreland Carolyn B. Oakley David K. Bwambok Vivian E. Fernand Narcisse

Background/Objectives: Over the past few decades, controlled substance abuse in drug-facilitated sexual assaults (DFSAs) has significantly increased worldwide, leading to an urgency to develop rapid and selective drug detection methods for field use (i.e., on-spot detection). Currently, techniques for detecting DFSA drug-associated samples are laborious and require skilled personnel to analyze/interpret the results. Moreover, most DFSA-associated drugs have a short half-life, making them more challenging to detect promptly. For instance, the timely detection of gamma-hydroxybutyrate (GHB) has been of ultimate concern for decades due to its fast elimination from the body. This study describes the development of a fluorescent ionic liquid nanosensor that can be used to rapidly detect GHB in the field. Methods: Trihexyltetradecylphosphonium fluorescein (THP2FL) ionic liquid was synthesized and evaluated for its potential application in detecting GHB. THP2FL nanoparticles in deionized water were synthesized with a size of 199 nm by a reprecipitation method. Results: The addition of GHB to THP2FL nanoparticles resulted in up to a 60% increase in fluorescence intensity and a 79% increase in absorbance. These results suggest potential applications for using the fluorescent THP2FL nanoparticles to detect GHB. The sensor’s selectivity was tested on compounds structurally similar to GHB, and the results showed that 1,4-butanediol (a precursor of GHB) is a potentially interfering species. Conclusion: This fluorescent technique allows for field deployable sensors, which would benefit screening GHB onsite.

Forensic Sciences doi: 10.3390/forensicsci5020027

Authors: Jonathan S. Lee-Confer Lila T. Wayman Kathryn L. Havens

Background/Objectives: Traditional diagnostic criteria for SBS include subdural, subarachnoid, and retinal hemorrhages. While intentional shaking is a known cause, the potential for similar forces acting on the head resulting from accidental trauma has not been fully explored. This study investigated the biomechanical forces on a model infant’s head during improper handling to determine if such forces could contribute to SBS without malicious intent. Methods: A realistic silicone infant model was equipped with an inertial measurement unit (IMU) to quantify head accelerations during two conditions: (1) placement of the infant model on a table with the head unsupported, and (2) manual shaking at maximum effort by 2 participants holding the model by the torso. Peak head accelerations were recorded for both conditions, and the results were analyzed for comparative assessment of the forces involved. Results: The average peak head acceleration when placing the infant model on a table with the head unsupported was +31,000 ± 7000 mg, with a range of +19,000.00 to +43,000 mg across trials. The average peak head accelerations during maximum effort shaking were significantly lower than placing the infant on the table, averaging 11,000 ± 10,000 vs. 31,000 ± 7000 mg, p < 0.0001). There were no significant differences in head accelerations between participants when placing the infant model on the table with the head unsupported (p = 0.89) nor with shaking the baby with maximum effort (p = 0.96). Conclusions: The study highlights that even accidental non-recommended handling of infants can result in high G-forces to the head, potentially leading to injuries similar to those observed in SBS. These findings highlight the necessity of supporting an infant’s head during handling and warrants caution against prematurely attributing physical abuse in SBS cases without considering unintentional causes.

Forensic Sciences doi: 10.3390/forensicsci5020026

Authors: Kuppareddi Balamurugan Elizabeth Staples Hussain Alghanim George Duncan Bruce McCord

Background: The identification of body fluids collected from crime scenes is crucial for determining the type and nature of assaults and for advancing the resolution of crimes. Objectives: The primary aim of this study was to investigate tissue-specific DNA methylation markers that can effectively distinguish buccal samples from blood, semen, and vaginal epithelial tissue. Methods: We screened various markers and selected four genomic locations for further analysis. Genomic DNA was extracted from tissue samples, followed by bisulfite conversion, locus-specific polymerase chain reaction (PCR) amplification, and pyrosequencing. Results: Four loci—cg-9652652, cg-11536474, cg-3867465, and cg-10122865—along with several adjacent CpG sites, were found to be hypermethylated in buccal samples compared to other tissue types. The difference in DNA methylation of buccal samples was statistically significant (p < 0.0001) compared to other tissues, indicating the potential usefulness of these loci for forensic tissue identification. Two additional studies were conducted: (a) a species specificity study and (b) a mixture study involving two different tissue types. The species specificity study showed that the primers used in the assay were specific to primates and humans. They did not amplify five non-primate samples, while the two primate samples—chimpanzee and rhesus—provided usable methylation data. The mixture study involved DNA from two different tissues—buccal samples and semen—combined in varying proportions. The results showed a decrease in the overall percentage of DNA methylation at the locus cg-9652652 as well as five adjacent CpG sites when the amount of buccal cell DNA in the mixture was reduced. Conclusion: The specificity of the primers and the significant differences in percent DNA methylation between buccal cells and other tissues make these markers excellent candidates for forensic tissue identification.

Forensic Sciences doi: 10.3390/forensicsci5020025

Authors: Samuel James Cockerill Emilio González-Reimers Matilde Arnay-De-La-Rosa

The calcaneus is very useful in archaeological contexts where human remains may be commingled. When DNA is impossible and the context is commingled, non-metric traits of the calcaneus may be used to identify phenotypical differences between human remains. Background/Objectives: We compared the prevalence of several calcaneal non-metric traits of pre-Hispanic and modern inhabitants of the Canary islands in order to (1) test the skeletal variation between pre-Hispanic and modern (17–18th century) Canarian samples and (2) gather information on biological and lifestyle differences between the samples using the calcaneal bone. Methods: We used a total of 364 calcanei (164 modern Canarian [72 left and 92 right] and 200 pre-Hispanic Canarian [92 left and 108 right] calcanei) and available non-metric traits to show differences between modern and pre-Hispanic Canarian population samples. Results: Our results highlight that some particular activity traits were common among the pre-Hispanic sample, while other traits, such as articular facet type Ib and the medial root of the inferior extensor retinaculum, show similar frequencies between pre-Hispanic and modern samples, which may indicate a genetic proponent influencing these similarities. Conclusions: Our results suggest that at least two traits, facet type Ib and the medial root of the inferior extensor retinaculum, may be influenced by genetics due to the persistence in modern samples despite the change of lifestyle between pre-Hispanic and modern Canarians.

Forensic Sciences doi: 10.3390/forensicsci5020024

Authors: Yangseung Jeong Aaron Gatewood Eun Jin Woo

(1) Background: Scavenger activity can pose significant challenges in forensic investigations. When examining damage on a victim, forensic investigators must first determine whether the damage resulted from human actions or animal scavenging. Accordingly, knowledge of the scavenger populations inhabiting a specific region and their scavenging behaviors is essential for accurately reconstructing the environmental context of the remains and interpreting associated taphonomic evidence in forensic investigations. (2) Methods: This study investigates the scavenging behavior of bobcats (Lynx rufus), a species native to North America, under controlled experimental conditions. The authors placed the carcasses of seven adult white-tailed deer in an outdoor field in Murfreesboro, Tennessee, and systematically documented the scavenging activities. (3) Results: Bobcats were observed to primarily scavenge soft tissue from the torso, focusing on musculature while generally avoiding bony regions. When unsatisfied with the torso, they shifted attention to the upper limbs, with minimal interest shown in the head or neck. Bobcats also exhibited a tendency to feed in a single area for extended periods, leading to concentrated damage in specific locations on the remains. (4) Conclusions: The findings demonstrate that bobcats can significantly influence the postmortem conditions of remains, producing characteristic patterns of modification. Recognizing these patterns is essential for forensic investigators and examiners when interpreting scavenger-related alterations in forensic contexts.

Forensic Sciences doi: 10.3390/forensicsci5020023

Authors: Dominic Gascho

The decision to remove a bullet from a gunshot victim depends on its location and associated medical risks, with surgical extraction often not indicated. Radiological imaging plays a vital role in assessing gunshot wounds and locating bullets, and it is essential in both clinical and forensic contexts. This narrative review examines the use of computed tomography (CT) for virtual bullet analysis, providing insights into shape, design, fragmentation, and material composition. Traditional 2D X-ray imaging, though commonly used, has limitations in accurately assessing caliber and position due to magnification and its 2D nature. In contrast, CT scans generate 3D reconstructions for detailed and precise examination, overcoming challenges such as metal artifacts with techniques such as extended Hounsfield unit (HU) reconstructions. These methods enhance the visualization of metal objects, allowing for better analyses of lodged bullets. Dual-energy CT further differentiates materials, such as lead and copper, using HU value differences at two energy levels. These advancements enable the virtual classification, shape analysis, and material identification of bullets in forensic investigations, even when the bullet remains in the body. As CT technology progresses, its forensic applications are expected to improve, providing more accurate and comprehensive differentiations of bullet types in future cases.

Forensic Sciences doi: 10.3390/forensicsci5020022

Authors: Cherrie Cheuk Yiu Chang Sys Stybe Johansen Brian Schou Rasmussen Kristian Linnet Ragnar Thomsen

Lithium is used as a medication in the treatment of bipolar disorder. Lithium has a narrow therapeutic index, and fatal intoxications have been described. The therapeutic drug monitoring of lithium is routinely performed in serum. Serum is commonly unavailable in forensic postmortem analysis, where whole blood is the matrix of choice. In this study, an inductively coupled plasma mass spectrometry (ICP-MS) method was developed and validated for the quantification of lithium in postmortem whole blood. Sample preparation consisted of a 100-fold dilution with acid and required only 40 µL of blood. Carry-over was deemed appropriately reduced with a rinse solution containing 5% hydrochloric acid. A nebulizer gas flow rate of 1.15 L/min showed a sufficient improvement of lithium sensitivity while simultaneously minimizing the background. Germanium was determined to be the most optimal internal standard. The method was validated in terms of linearity, accuracy, precision, and lower limit of quantification. Linearity was demonstrated within the analytical measurement range of 0.10–1.5 mmol/L. The method showed acceptable precision and accuracy, with a total coefficient of a variation ≤2.3% and accuracies ranging from 105 to 108% at all concentrations in the quality control samples. The final method was applied to postmortem blood from 103 consecutive autopsy cases and demonstrated robustness by low intermediate precision and high and consistent recovery of the internal standard.

Forensic Sciences doi: 10.3390/forensicsci5020021

Authors: Ana Zoe Monogan Joshua L. Smith Paola A. Prada-Tiedemann

Background: Chemical odor profiling within forensic entomology is an emerging tool given its potential for species identification and larval aging and its ability to identify decomposition stages. A volatile analysis of larval masses across species of distinctive developmental stages was carried out with extraction techniques to identify odor signatures. However, it is unknown how larval sample handling (i.e., live samples in research vs. hot-water-killed samples in casework) affects odor signatures or the possibility of obtaining relevant volatiles from a single larva. Method: This study utilized solid-phase microextraction (SPME) with gas chromatography–mass spectrometry (GC/MS) for the analysis of odor volatiles emanating from single larval samples of Cochliomyia macellaria. Fifty (50) larvae (25 live; 25 boiled) were analyzed. Results: The SPME-GC/MS method allowed for odor volatile detection from a single maggot regardless of the sample handling group. The main compounds identified across both groups included those previously reported as emanating from larvae and decomposition substrates. When comparing treatments, the boiled larval samples had a 6-fold decrease in compound abundance compared to the live samples. The identified odor volatiles observed in the hot-water-killed treatment group included indole, p-cresol, and phenol. Conclusions: These results suggest that the handling technique impacts odor detection. Additionally, the heterocyclic aromatics and alcohols identified in the boiled samples are potentially odor markers of a higher intrinsic nature to the maggot rather than a cross-transfer from the decomposition substrate given their survival post elevated temperature treatment. This work shows the plausibility of carrying out an odor analysis of a single maggot following both common research and casework handling practices.

Forensic Sciences doi: 10.3390/forensicsci5020020

Authors: Mario Chisari Martina Francaviglia Sabrina Franco Gianpietro Volonnino Raffaella Rinaldi Nicola Di Fazio Lucio Di Mauro

Background: The COVID-19 pandemic significantly disrupted forensic science, exposing vulnerabilities and introducing unprecedented challenges. Five years later, its impact persists, necessitating ongoing adaptations in forensic practice. This study examines key transformations, persistent issues, and emerging challenges in forensic science post-pandemic. Methods: A critical analysis of forensic science’s response to the pandemic was conducted, focusing on operational disruptions, methodological advancements, educational shifts, and technological integration. Results: Forensic operations faced delays due to case backlogs, restricted in-person work, and postponed court proceedings. Forensic pathology evolved with increased reliance on molecular autopsy techniques to clarify COVID-19-related deaths. Educational methods shifted toward virtual learning, prompting discussions on standardized digital training. Additionally, artificial intelligence and automation gained prominence in forensic investigations, enhancing crime scene analysis and predictive modeling. Discussion: While forensic science demonstrated adaptability, challenges remain in international collaboration, resource distribution, and professional training. The pandemic accelerated technological integration but also raised ethical and procedural concerns, particularly regarding AI applications in legal contexts. Virtual learning innovations necessitate further development to ensure competency in forensic training. Conclusions: Forensic science continues to evolve in response to post-pandemic realities. Addressing gaps in cooperation, technology implementation, and training will be crucial to strengthening the field. By assessing these changes, this study underscores forensic science’s resilience and adaptability, offering insights into its future trajectory amid ongoing challenges.