Forensic Sciences doi: 10.3390/forensicsci6030071

Authors: Massimiliano Esposito Federica Ministeri Mario Giuseppe Chisari Lucio Di Mauro Serena Matera Valentina Schilirò Ermanno Vitale Francesco Sessa Giuseppe Ragazzi Carmelo Ferrara Cristoforo Pomara

Background: Health assessment in custodial settings represents a core medico-legal function, particularly when clinical changes may influence detention compatibility and judicial decisions. Objective documentation of disease progression during incarceration is essential to ensure respect for the principle of healthcare equivalence and the protection of fundamental rights. Methods: A retrospective descriptive medico-legal analysis was conducted on 37 male inmates referred for forensic evaluation. Standardized assessments included anamnesis, physical and psychiatric examination, evaluation of medical records, and comparative evaluation of pre-detention and detention-period health status. Results: Pre-existing conditions accounted for 73% of pathologies, while 27% developed during imprisonment. Clinical deterioration occurred in 35.1% of inmates (13 cases). Psychiatric disorders were frequent (27%, 10 inmates), predominantly depressive. Conclusions: The findings indicate that imprisonment may constitute a setting that contributes to the progression of medical and psychiatric conditions, highlighting structural and organizational challenges within penitentiary healthcare. Strengthening standardized medico-legal assessment protocols, improving multidisciplinary collaboration, and ensuring continuity of care between prison and community health services are crucial steps toward protecting inmates’ health. This study provides an exploratory assessment of health conditions in a cohort of inmates within the Italian correctional system, emphasizing the central role of healthcare in custodial settings. Since many Italian correctional facilities are equipped with well-developed healthcare services and appropriate diagnostic and therapeutic resources, numerous acute and chronic medical conditions can be effectively managed within the prison setting, ensuring continuity of care while limiting the need for transfer to external healthcare facilities.

Forensic Sciences doi: 10.3390/forensicsci6030070

Authors: Anna Barbaro Angelo La Marca Giacomo Falcone

Background: The Investigator Quantiplex Pro RGQ Kit (Qiagen, Hilden, Germany) enables rapid quantification of total human and male DNA on the Rotor-Gene Q platform while simultaneously assessing PCR inhibition, DNA degradation, and mixed DNA profiles. This study evaluated whether reducing the recommended qPCR reaction volume by half affects analytical performance, with the aim of improving throughput and reducing reagent costs in forensic laboratories. Methods: Control DNA (50 ng/µL) and DNA extracted from 20 simulated forensic samples representing common forensic evidence (including bloodstains, semen stains, saliva samples, vaginal swabs, touch DNA, hair roots, and mixed biological traces deposited on different substrates) were analyzed using standard and half-volume qPCR reactions with identical reagent proportions. Quantification and quality indices—including inhibition (IC), degradation (DI), and mixture (MI)—were evaluated using Q-Rex software. Ct values, ΔCt, and standard deviations were compared between protocols using paired statistical analysis, with significance set at p < 0.05. Results: Standard curves generated under both reaction conditions showed efficient amplification, with R2 values > 0.998 and slopes within recommended ranges. Control samples demonstrated strong agreement between expected and measured DNA concentrations for both protocols. Although forensic samples displayed variable DNA quantity and quality, highly concordant results were obtained between standard and reduced volumes. Mean Ct values showed minimal ΔCt variation and low dispersion across all sample types. IC, DI, and MI indices were also consistent, indicating reliable detection of inhibition, degradation, and mixed DNA profiles under reduced-volume conditions. Conclusions: Half-volume qPCR reactions using the Investigator Quantiplex Pro RGQ Kit maintained accurate, precise, and reproducible DNA quantification and quality assessment. Reduced-volume protocols may therefore provide a cost-effective strategy to increase throughput and improve workflow efficiency in forensic laboratories without compromising analytical robustness.

Forensic Sciences doi: 10.3390/forensicsci6030069

Authors: Csongor Herke Vladimir Aleksandrovich Fedorenko

Background/Objectives: This paper focusses on the use of artificial intelligence (AI) in automated firearm and ammunition identification. It concentrates on firing pin impressions on fired cartridge cases and secondary rifling land impressions on fired bullets, since these traces represent two different problems in forensic ballistic comparison. The main question was whether machine learning and deep learning methods can support the comparison process while the final assessment remains in the hands of the firearms examiner. Methods: This study combines a critical methodological analysis of AI-based firearm identification with applied experimental results. A convolutional neural network (CNN) was used for firing pin mark classification, the reliability of which was assessed using output neuron parameters A1, A1/A2, and A1−A2. For bullet marks, CNN-based semantic binarization was applied to secondary rifling land impressions, followed by random forest classification of the binarized image pairs. Data augmentation was used to address the limited number of original training objects in this study. Results: Considering the three highest CNN output signals increased the overall firing pin classification accuracy from 82.6% to 92.8%, while the accuracy for unknown-class marks increased from 60.8% to 79.1%. CNN-based binarization of bullet marks achieved accuracy = 0.88 ± 0.05, recall = 0.76 ± 0.06, precision = 0.83 ± 0.06, F1 = 0.79 ± 0.05, and MCC = 0.71 ± 0.06. The random forest classification of binarized bullet mark pairs achieved an accuracy of approximately 84–86%. Conclusions: The findings indicate that AI can improve speed, consistency, candidate selection, image preprocessing, and reliability assessment in forensic ballistics. However, AI outputs remain dependent on dataset quality, firearm and ammunition variability, unknown class recognition, and external validation. The most reliable model is a hybrid human–AI workflow in which the algorithm supports the firearms examiner but does not replace expert judgement.

Forensic Sciences doi: 10.3390/forensicsci6030068

Authors: Mmabatho Portia Aphane Jacob Tseko Mofokeng

Background/Objectives: Cybercrime poses a growing threat to South Africa’s digital economy, necessitating effective search and seizure practices for digital evidence; however, investigators and prosecutors encounter systemic challenges that hinder successful prosecutions. This study aims to identify and analyse these obstacles and propose improvements to enhance the efficiency and integrity of digital forensic processes within the criminal justice system. Methods: This empirical study employed a qualitative research design, collecting data from 30 participants within the South African Police Service (SAPS) Cyber-Crime and Deep Web Investigation Unit and the Specialised Commercial Crime Unit (SCCU); semi-structured interviews were conducted and analysed thematically to identify recurring patterns and institutional challenges. Results: The findings reveal that digital investigations are complex, multi-stage processes requiring a strictly auditable approach to preserve the integrity of evidence from seizure to courtroom presentation; a documented chain of custody is critical to ensuring that digital evidence remains in its original, unaltered state, while institutional silos disrupt the effective flow of evidence to prosecutors despite the use of specialised forensic methodologies. Conclusions: The study concludes that addressing these challenges requires the establishment of a multi-agency digital forensic task force and specialised interdisciplinary training to bridge the technical-legal gap, with significant implications for policy reform and the advancement of digital forensic practices in South Africa.

Forensic Sciences doi: 10.3390/forensicsci6030067

Authors: Elena Sierra-Serrano John Albanese

Background/Objectives: Age estimation from skeletal remains is an important step in reconstructing the identity of deceased individuals. The auricular surface of the ilium, known to be a strong predictor of age at death, could benefit from methodological refinements that have previously shown potential using only the retroauricular area. The aim of this paper is to showcase the strengths and weaknesses of an approach that requires greater complexity than using only the retroauricular area. Methods: A series of logistic regressions were performed to classify 105 individuals from the Terry Collection into three age groups: younger, middle-aged, and older adult. Stratified sample selection provided a more all-encompassing reflection of the diverse experiences of those individuals who carried the morphologies observed. The features scored included microporosity, macroporosity, presence of dense bone, morphology of the auricular rim, linear organization, and retroauricular area. Results: Overall, the substantial increase in time and effort does increase accuracy of the classifications, but not by the same magnitude. Results reflect an improved classification of younger individuals compared to using the retroauricular area alone. With the present method, individuals were classified into their correct group 66% of the time, and results that narrowed down classification to two of the three age groups were obtained in a further 26%. Conclusions: The utility of this method is based on clarity and reproducibility, addressing the known issue of “attraction to the middle”, and its potential to correctly identify adults as beyond 65 years old.

Forensic Sciences doi: 10.3390/forensicsci6030066

Authors: Beatrice Benedetti Vincenzo M. Grassi Caterina Pesaresi Fabio De-Giorgio

Background: Death following blunt force trauma in the absence of major internal injuries represents a significant forensic challenge. In such cases, reflex-mediated mechanisms leading to cardiac inhibition may be overlooked, particularly when multiple autonomic triggers act simultaneously. Methods: We report the case of a 78-year-old man who died after a violent assault involving repeated blunt force trauma, followed by a prolonged 16-day submersion in river water. A complete forensic investigation was performed, including postmortem computed tomography, autopsy, histological examination, toxicological analyses, and diatom testing to assess the possibility of drowning. Results: External and internal examination revealed multiple blunt force injuries involving the head, face, thorax, upper limbs, scrotal region, and back. Rib fractures, pulmonary contusions, and testicular hematomas were documented, but no internal lesion of sufficient severity to independently explain death was identified. No significant pre-existing cardiac disease or structural cardiac abnormality was found. Toxicological analysis showed a blood alcohol concentration of 0.7 g/L, with no lethal substances detected, although interpretation was limited by the prolonged 16-day submersion interval. Diatom testing performed on multiple tissue samples was negative, arguing against drowning. Overall, neither trauma, intoxication, nor submersion alone adequately explained the fatal outcome. Conclusions: This case supports the hypothesis that parasympathetic hyperactivation may have contributed to death. The combined effects of trauma to reflexogenic areas, cold water immersion, and alcohol-related autonomic modulation may have triggered a fatal cardioinhibitory response. Reflex-mediated death should be carefully considered in similar cases when conventional autopsy findings are inconclusive. In the present case, interpretation is limited by prolonged submersion and related postmortem changes, which may have affected the reliability of certain forensic findings.

Forensic Sciences doi: 10.3390/forensicsci6030065

Authors: Mpho Maboa

Background/Objectives: The integrity of firearm evidence is central to solving violent crimes and securing just court outcomes, yet the consistency of investigative practices across policing environments remains uncertain. This study examined the articulated understanding and reported practices related to firearm evidence among investigating officers, crime scene experts from the Local Criminal Record Centre (LCRC), and ballistics experts within the South African Police Service (SAPS). Materials and Methods: A qualitative research design was adopted, using semi-structured interviews with 39 participants across the three groups. The study explored participants’ knowledge of firearm evidence, types of evidence found at murder crime scenes, procedures for evidence collection, and the concept of chain of custody. Data was analysed thematically to identify patterns and knowledge gaps. Results: The findings revealed considerable variation across groups in articulated knowledge and reported procedural awareness, with crime scene experts demonstrating stronger conceptual clarity, while investigating officers showed notable gaps in recognising trace elements and maintaining continuity of possession. Ballistics experts possessed technical expertise but limited familiarity with scene-level protocols. These disparities have the potential to pose risks to evidentiary integrity and evidentiary admissibility if reflected in practice. Conclusions: The findings suggest that enhancing shared understanding of firearm evidence handling through reinforced procedural guidance and professional development may support greater consistency at early investigative stages. These conclusions are grounded in participants’ articulated knowledge and reported procedural awareness derived from interview data, rather than observed investigative practice or case file compliance. Further research is required to assess how such interventions influence operational practice and to explore the potential role of digital tools in supporting chain-of-custody continuity.

Forensic Sciences doi: 10.3390/forensicsci6030064

Authors: Mpho Maboa

Background/Objectives: Despite its central role in murder investigations, firearm evidence remains one of the most scientifically contested and legally scrutinised forms of forensic proof. International critiques have highlighted limitations in foundational validity, examiner subjectivity, and individualisation claims within firearm and toolmark analysis, underscoring the importance of examining these issues within the South African context. Despite its central role in murder investigations, firearm evidence remains one of the most scientifically contested and legally scrutinised forms of forensic proof. Firearm evidence remains central to murder investigations; however, ongoing scientific and legal debates continue to raise concerns about its reliability, interpretation, and admissibility. Materials and Methods: A qualitative case study approach was used to examine the scientific basis, forensic integrity, and legal admissibility of firearm evidence. Semi-structured interviews were conducted with 39 purposively selected participants, including detectives, crime scene experts, and ballistics specialists. Participants were drawn from Tshwane-based police stations with high murder rates, using a combination of purposive, random, and systematic sampling techniques. Guided by interpretivist and constructivist paradigms, thematic and content analysis were applied building inductively on the empirical findings and informed by contemporary international debates. The study developed the Triadic Model of Forensic Investigation: Science, Integrity, and Law, integrating scientific, procedural, and legal dimensions of firearm evidence. Results: The study examined the Locard Exchange Principle as the scientific foundation of firearm evidence and found varying levels of practitioner understanding. Although some participants demonstrated foundational knowledge of forensic concepts such as identification and individualisation, terminology and conceptual understanding varied substantially across professional groups. Understanding of chain-of-custody requirements and legal admissibility criteria was uneven, with notable gaps among some practitioner groups. These inconsistencies, particularly among Investigating Officers, have significant implications for evidentiary integrity and courtroom reliability, especially in contexts where courts increasingly scrutinise categorical claims of firearm source attribution. Conclusions: The results highlight the importance of aligning professional practice, forensic education, and evolving scientific and legal standards to support reliable and legally admissible firearm evidence. The Triadic Model provides a coherent evaluative structure for strengthening the reliability, interpretive accuracy, and legal defensibility of firearm evidence. The study emphasises the value of interdisciplinary collaboration and closer alignment between forensic science, operational practice, and evolving judicial standards.

Forensic Sciences doi: 10.3390/forensicsci6030063

Authors: Julia Meyers Lesley Harrington Deborah C. Merrett Hugo F. V. Cardoso

Background/Objective: Most weight estimation formulae developed for children from skeletal measurements are based on 20 individuals from the Denver Growth Study. This limitation is mainly due to the availability of data. The formulae produced using these data are limited to a small range of variation in weight, which may result in reduced applicability. This study develops new weight estimation formulae relying on a larger and more diverse sample of deceased children (n = 77) of known age and weight from the New Mexico Decedent Image Database. Methods: A classical calibration regression approach was used to generate weight estimation equations from virtual metaphyseal breadths, epiphyseal breadths, and torsional rigidity (J) of the femur and tibia. Equations were developed separately for the two sexes, two BMI categories, and three age categories. Results: Midshaft femoral J values (45.5% of the total diaphyseal length) and proximal tibial J values (75% of the total diaphyseal length) correlate most highly with body weight and when used to develop weight estimation formulae produce the least amount of error. In general, the breadth measures were less correlated and had larger error than the J values, though the distal femoral metaphyseal breadth produced the highest coefficients of determination and lowest error of the breadth formulae. Conclusions: While these formulae are based on a sample of modern children, the variation that they encompass and the different equations provided allow users to apply them in a variety of circumstances. However, weight estimation may have limited value due to large prediction errors.

Forensic Sciences doi: 10.3390/forensicsci6030062

Authors: Emanuele Vicario Fabio Comero Gabriele Gaggero Francesco Ventura Rosario Barranco

Background and Clinical Significance: Necrotizing fasciitis (NF) is a fulminant, life-threatening polymicrobial infection of the soft tissues, characterized by the progressive and widespread necrosis of the subcutaneous tissues and fascial planes. Although the cervical region is an infrequent site of involvement, cervical necrotizing fasciitis (CNF) is distinguished by its rapid anatomical spread, severe systemic complications, and significant mortality rates. Case Presentation: This report details the fatal clinical course of a 37-year-old immunocompetent male with no relevant active comorbidities. The patient initially presented with insidious, non-specific symptoms clinically interpreted as an uncomplicated upper respiratory tract infection/acute pharyngitis, including pharyngodynia, hoarseness, fever, lateral cervical lymphadenopathy, and catarrhal secretion. Within a brief period, his clinical status underwent rapid deterioration, with progressive cervical swelling, dysphonia, swallowing impairment, dyspnoea, shock, and subsequent multi-organ dysfunction syndrome. A total-body computed tomography scan revealed lateral cervical subcutaneous emphysema, a small pneumomediastinum, bilateral pleural effusion and bilateral pulmonary parenchymal consolidations. Conclusions: A comprehensive forensic examination revealed swelling and brownish-green discoloration of the right cervical region, palpable crepitus, and extensive necrosis of the cervical soft tissues, with involvement of the deep fascial layers and regional musculature. Histological examination confirmed massive cervical soft tissue necrosis associated with inflammatory infiltration and air bubbles. Multifocal acute pneumonia was also documented. This case underscores the importance of a meticulous examination of the cervical region during autopsy in suspected infection-related fatalities. Clinically, it highlights the diagnostic challenges posed by the deceptive and paucisymptomatic onset of CNF.

Forensic Sciences doi: 10.3390/forensicsci6030061

Authors: Raisa Agustín María José Miñano Noèlia Ortuño Jesus Cobo Diego J. Palao

Background: Competence (within the legal and bioethical framework in Spain, the term ‘competence’ is used as equivalent to healthcare decision-making capacity) in the field of child and adolescent psychiatry is a complex and evolving issue. Legal frameworks vary by country and legal tradition. Most concerns relate to treatment decisions within mental health, but in fact there is a wide range of decisions with a relevant impact on patients’ lives and quality of life over the long term. Forensic science must adapt its current practice to the worldwide challenge presented by this evidence. Objectives: This article will present a current research project about competence (healthcare decision-making capacity) in a sample of adolescent and child population in Spain. The final objective will be to validate the Spanish version of the MacCAT semi-structured interview consenting to psychiatric admission in adolescents in a mixed sample. We also examine some of the influences that must be taken into account in this research. Methodology: This is an instrumental and methodological study utilizing a sequential mixed-methods design (qualitative content validation followed by a quantitative psychometric evaluation). A process of adaptation of the Spanish MacCAT to consent to psychiatric admission in the adolescent population. A pilot study will initially be conducted to evaluate question comprehension and duration. Subsequently, a cross-sectional study will be carried out to assess inter-rater reliability, internal consistency, and validity. Evaluation will include clinical and neuropsychological assessment in five departments at two University Hospitals. Expected results: We will establish new evidence about the evaluation of competence in adolescent psychiatry; to date, no studies have been identified assessing the capacity to consent to psychiatric admission among Spanish-speaking adolescents, nor the factors potentially related to such competence. To our knowledge, the relationship between competence and illness awareness in the adolescent population remains unexplored in the literature, and its analysis constitutes one of the novel contributions of the present study. In addition, we will adapt and validate a well-known evaluation scale to the specific environments of our medical and psychiatric health departments. Conclusions: Objective evaluation of competence for medical interventions and treatments is a challenge in clinical and forensic examination. New evidence is necessary to operationalize classical evaluations in standardized ways, adapted to different countries, legal systems and forensic traditions.

Forensic Sciences doi: 10.3390/forensicsci6030060

Authors: Martina Di Santo Paolo Fais Lina De Paola

Background: Recent technological advances, such as 3D laser scanning, digital photogrammetry and virtual autopsy, have significantly reshaped the acquisition and interpretation of scientific evidence in criminal proceedings. These tools enable highly detailed visualizations that can enhance factual understanding while also introducing potential cognitive biases for judges. This study aims to assess the legal, epistemological and forensic implications of 3D virtual reconstruction as evidence, examining its admissibility, scientific robustness, and associated risks within contemporary criminal trials. Methods: An interdisciplinary methodology was applied, integrating criminal procedural law, forensic medicine, digital forensics and legal epistemology. The study includes doctrinal analysis, a review of the literature from the last decade, Italian and comparative case law, and evaluation of European forensic guidelines (particularly ENFSI). Direct observation of laboratory practices and technical assessment of reconstruction tools were conducted to evaluate scientific validity, repeatability, traceability and transparency. Results: The findings reveal that 3D reconstruction provides high metric accuracy, durable digital preservation of the crime scene and improved communication of complex dynamics to the court. It allows experts to test alternative hypotheses, reducing ambiguity in technical explanations. Nevertheless, several critical issues emerged: the persuasive power of highly realistic imagery, dependence on non-transparent software processes, potential vulnerabilities in the chain of custody for digital data, and the absence of a unified Italian regulatory framework governing digital scientific evidence. Conclusions: 3D virtual reconstruction constitutes a powerful but epistemologically complex form of scientific evidence. Its probative value depends on transparent methodology, verifiability and strict compliance with adversarial safeguards. The study underscores the need for national technical standards, clear admissibility criteria and specialized training for legal professionals. A coherent regulatory framework is essential to ensure that digital technologies enhance, rather than distort, the pursuit of truth in criminal justice.

Forensic Sciences doi: 10.3390/forensicsci6030059

Authors: Oshoke Samson Igonor Muhammad Bilal Amin Saurabh Garg

Background/Objectives: The rapid expansion of digital evidence in modern investigations has created pressing challenges for maintaining integrity, traceability, chain of custody, and meaningful analysis across heterogeneous forensic artefacts. Conventional evidence management approaches often fall short in scalability, transparency, and the ability to correlate diverse digital evidence. This study presents BlockFECS, a blockchain-based proof-of-concept system for metadata-driven evidence correlation in digital forensics. Methods: BlockFECS uses Hyperledger Fabric to support auditable and tamper-resistant evidence management while capturing structured forensic metadata, including timestamps, locations, device IDs, user IDs, and file hashes. An off-chain weighted correlation algorithm assigns similarity scores between evidence pairs and classifies relationships as Related, Supplementary, Duplicate, or Unrelated. The system was evaluated using a simulated smart city accident scenario and tested for correctness, transaction latency, throughput, scalability trends, and concurrency behaviour across four computing environments. Results: Within the controlled proof-of-concept dataset, the correlation algorithm achieved 1.00 precision and recall for clear Related and Duplicate evidence relationships and high precision (0.90) for Supplementary relationships, although recall in this category was lower due to incomplete or noisy metadata. Performance testing showed that Create, Transfer, and Delete operations completed with sub-second latency, while correlation throughput exceeded 60 comparisons per second across all tested environments. Conclusions: The findings demonstrate the feasibility of combining blockchain-backed evidence integrity with lightweight metadata-driven forensic intelligence. BlockFECS contributes a proof-of-concept model for automating metadata-based evidence analysis while preserving provenance integrity and auditability, highlighting a promising direction for trustworthy and intelligent digital forensic investigation support.

Forensic Sciences doi: 10.3390/forensicsci6030058

Authors: Vincenzo Cianci Anna Messina Maria Manno Daniela Sapienza Desirèe Speranza Ludovica Pepe Vincenzo Fiorentino Patrizia Gualniera Alessio Asmundo Cristina Mondello

Introduction: Otorrhagia is an uncommon finding in forensic practice and is classically associated with skull base fractures, temporal bone trauma, or barotraumatic events. However, the literature suggests that ear bleeding may also rarely occur in deaths due to neck compression, including strangulation and hanging. Case Presentation: We report the case of a man died by hanging. External examination revealed an oblique ligature mark on the neck, conjunctival petechiae, and bilateral otorrhagia. At autopsy, no fractures of the skull vault or skull base were identified. Otoscopic examination showed bilateral tympanic membrane laceration. Internal examination of the neck disclosed small hemorrhagic infiltrates in the perithyroid muscles and prevertebral soft tissues, as well as bilateral carotid intimal tears consistent with Amussat’s sign. Moreover, a unilateral lesion involving the petrous portion of the temporal bone was observed. The distinctive feature of this case is the association of bilateral otorrhagia, bilateral tympanic membrane laceration, and unilateral petrous bone erosion in the absence of skull base fracture. Histology showed visceral congestion, pulmonary overdistension with rupture of interalveolar septa, cerebral edema, myocardial fiber fragmentation and trabecular remodeling and areas of necrosis of the bone fragment sampled from the right petrous temporal bone. Death was attributed to mechanical asphyxia due to hanging. Conclusions: This case suggests that bilateral otorrhagia in hanging may result from the combined effects of impaired cervico-cephalic venous drainage and pressure changes transmitted to the middle ear during forceful respiratory efforts. The pre-existing unilateral petrous bone lesion may have acted as a local predisposing factor, although its etiology and actual contribution cannot be established with certainty. Otoscopic examination should therefore be regarded as a potentially useful adjunct in selected cases, particularly for documenting auricular findings and supporting differential diagnosis.

Forensic Sciences doi: 10.3390/forensicsci6030057

Authors: Amisha Nayak Sandhya Tamgadge Junaid Ahmed Srikant Natarajan Nandita Shenoy Pradeep Sherigar Nanditha Sujir

Background/Objectives: Nonmetric dental traits assessed using the Arizona State University Dental Anthropology System (ASUDAS) are valuable in forensic identification due to their population-specific variation. However, intranational variability within Indian populations remains underexplored. To evaluate variation in ASUDAS dental morphological traits among individuals with different early-life geographic backgrounds and assess their forensic applicability. Methods: A cross-sectional observational pilot study was conducted on 55 dental casts of individuals aged 18–22 years. Subjects were grouped into Maharashtra (n = 37) and non-Maharashtra (n = 18) based on residence from birth to 10 years. A total of 42 crown traits were assessed using ASUDAS criteria. Statistical analysis included chi-square or Fisher’s exact test (p < 0.05), and intraobserver reliability was evaluated using Cohen’s kappa. Results: Significant differences were observed in maxillary traits such as shoveling (p = 0.004), interruption grooves (p = 0.01), canine accessory distal ridge (p = 0.022), hypocone (p = 0.029), premolar accessory ridge (p = 0.007), tuberculum dentale (p = 0.021), and double shoveling (p = 0.001), and mandibular traits including premolar accessory cusp/protoconule (p < 0.001), anterior fovea (p = 0.005), and deflecting wrinkle (p < 0.001). Conclusions: The observed variations reflected population heterogeneity, supporting the forensic relevance of ASUDAS traits and the need for region-specific databases.

Forensic Sciences doi: 10.3390/forensicsci6030056

Authors: Isabel Almeida Pedro M. Teixeira José Manuel Teixeira Teresa Magalhães

Background/Objectives: Health-related quality of life perception (HRQoL) reflects the impact of individuals’ health conditions on their physical, psychological, and social well-being, and can be compromised after an accident The general aim of this study was to analyze the effect of occupational accident (OA) outcomes on injured workers’ HRQoL. Methods: We conducted a cross-sectional study, using a convenience sample of 101 participants at the end of their recovery and professional reintegration (PR) process. They were submitted to a personal injury assessment (PIA) conducted by medico-legal specialists, and data related to injury severity (IS), permanent professional disability (PD), and PR were collected from the respective forensic reports. Subsequently, they underwent a psychological interview and filled out self-report questionnaires to measure HRQoL (SF-36) and resilience (RSA). For each variable, two groups were defined. Analyses included descriptive statistics, correlations, group comparisons, and multiple linear regression analyses. Results: Injured workers reported lower HRQoL than Portuguese norms across most domains. HRQoL scores were positively associated with resilience and PR, and negatively associated with IS and PD. In multivariable models, IS, and RSA emerged as significant independent associated variables of the physical–social HRQoL component. Conclusions: These findings highlight the importance of a biopsychosocial and multidisciplinary approach to OA victims’ professional reintegration, integrating physical treatment and psychological support with resilience-building and work rehabilitation, before medical discharge and PIA.

Forensic Sciences doi: 10.3390/forensicsci6030055

Authors: Tomáš Vokálek Martin Drahanský

Background/Objectives: Latent fingerprints are routinely recovered from conventional porous and non-porous substrates; however, biologically active surfaces such as plant leaves are generally regarded as unsuitable for dactyloscopic evidence. Because vegetation is frequently present at crime scenes, this study aimed to systematically evaluate whether plant leaves can retain usable friction ridge detail and to determine the durability and forensic value of such traces under laboratory and outdoor conditions. Methods: Latent fingerprints were deposited on leaves of multiple plant species (maple, ash, dandelion, bird cherry, chestnut, climbing ivy, and five-leaved ivy) under dry and hydrated conditions and at defined time intervals after deposition. Visualization was performed using several powders, with SupraNano Fluorescent Green magnetic powder providing the best performance. Developed impressions were photographed using controlled illumination and evaluated using automated quality assessment (NFIQ 2.0) and comparison software (Innovatrics IDkit 9.1.7.1004). Additional experiments examined living, growing leaves exposed to natural weather conditions for extended periods. Results: Usable ridge detail was successfully visualized on all tested species. Bottom leaf surfaces and hydrated samples generally provided better preservation and contrast. Identifiable traces persisted for up to 20 h on detached leaves and for up to 35 days on living leaves despite growth-related deformation. Under outdoor exposure, fingerprints on ivy remained visible and comparable for up to 60 days. Although overall automated quality scores were reduced by background venation, selected impressions achieved measurable comparison scores and successful matches. Conclusions: Plant leaves can serve as unconventional yet viable carriers of latent fingerprints. Magnetic fluorescent powder development combined with careful documentation enables recovery of forensically useful ridge detail even after prolonged environmental exposure. These findings expand the range of substrates that should be considered during crime scene processing and provide practical guidance for evidence collection on vegetation.

Forensic Sciences doi: 10.3390/forensicsci6020054

Authors: Ștefania Andrei Daria Hemeș Adelina Dubaș Alina Frunză Daniela Gabriela Andrei Cosmina Chivu Anna Florescu Matei Dragodan Albert Merfu Bogdan Pădure Sorin Hostiuc

Background/Objectives: Child sexual abuse (CSA) is a serious public health and forensic issue, associated with significant psychological, psychiatric, and social consequences. However, data from forensic psychiatric evaluations regarding victims of this type of abuse are limited, especially in Romania. This study primarily aimed to profile the demographic, clinical, and psychosocial characteristics of minors who were victims of sexual offenses and underwent forensic psychiatric evaluation. Secondary objectives included examining relationships between demographic and clinical variables, describing patterns of victimization, and analyzing the co-occurrence of vulnerability factors. Methods: This study was retrospective, descriptive, and analytical, including 51 minors (under 18 years of age) examined at the Mina Minovici National Institute of Legal Medicine (INML), between 2013 and 2025. Data extracted from forensic psychiatric reports included demographic variables, the type of offense, psychiatric diagnosis, psychological issues, intellectual functioning, family environment, and relationship with the aggressor. Statistical analyses included descriptive statistics, Pearson χ2 and Fisher’s exact tests, binary logistic regression, and nonparametric tests for cumulative vulnerability. Results: The sample was predominantly female (94.1%), with a mean age of 13.75 years. Rape was the most frequent offense (45.1%). Most victims originated from disorganized family environments or were institutionalized (70.6%), and 60% had below-normal intellectual functioning. A psychiatric diagnosis was present in 56.9% of cases, while 70.6% exhibited psychological issues. Within an exploratory logistic regression model that did not reach overall statistical significance (omnibus likelihood ratio test χ2 = 9.31; p = 0.097), the presence of a psychiatric diagnosis showed the strongest individual association with psychological issues (OR = 5.17; 95% CI, 1.22–21.85; p = 0.026). Cases in which the aggressor was not related to the family environment were the most frequent (60.8%), followed by cases involving family members (23.5%). Most subjects displayed multiple co-occurring vulnerability factors. When the cumulative vulnerability score was recalculated to exclude psychological issues (to avoid construction-induced circularity), higher vulnerability remained significantly associated with extrafamilial victimization (Kruskal–Wallis H = 6.93; p = 0.031), but was no longer associated with psychological issues themselves (Mann–Whitney U = 298.0; p = 0.56), indicating that the originally observed association was an artefact of the score’s construction. Conclusions: Minors who are victims of sexual abuse often face multiple overlapping vulnerabilities, including unstable family environments, cognitive impairments, and psychiatric conditions. These factors frequently coexist in victims evaluated in forensic psychiatric settings and are associated with psychological issues and complex victimization profiles. The findings emphasize the importance of comprehensive, multidisciplinary, and victim-centered methods in forensic psychiatric evaluations.

Forensic Sciences doi: 10.3390/forensicsci6020053

Authors: Athip Lorsirigool Narong Kulnides Natapol Pumipuntu Pratch Sukumolanan Benjamabhorn Pumhirunroj

Background and Clinical Significance: Animal hoarding is a recognized form of animal cruelty characterized by the accumulation of animals under conditions of inadequate care, leading to compromised welfare and an increased risk of disease transmission. Veterinarians are often the first to identify such cases through clinical presentation. This case report describes a forensic investigation of a dog associated with animal hoarding and environmental neglect in Bangkok, Thailand. Case Presentation: A 7-year-old male mixed-breed dog was presented with seizures, emaciation, dehydration, and ectoparasite infestation. Hematological findings revealed leukopenia and anemia, and diagnostic testing confirmed infection with canine distemper virus (CDV) and Dirofilaria immitis. Despite supportive treatment, the dog died within 48 h. A subsequent site inspection, conducted using the TRACE model and based on crime scene investigation principles, identified 13 dogs housed in overcrowded and unsanitary conditions. Environmental assessment revealed poor sanitation, waste accumulation, and fluorescent stains of unknown origin detected under ultraviolet light (365 nm) in the cage areas. Among the animals, infections with CDV (23.08%), Dirofilaria immitis (53.85%), and Ehrlichia canis (61.54%) were identified, indicating a high burden of infectious and vector-borne diseases. Conclusions: This case demonstrates the value of integrating clinical veterinary findings with forensic investigation in the assessment of animal neglect. The combined use of clinical, laboratory, and environmental evidence strengthens documentation and supports potential legal action. Enhanced veterinary forensic training and standardized protocols are recommended to improve early detection and intervention in animal cruelty cases.

Forensic Sciences doi: 10.3390/forensicsci6020052

Authors: Pietro Zuccarello Giulia Carnazza Massimiliano Esposito Grazia Giulia Pantè Martina Francaviglia Francesco Sessa Nunziata Barbera

Background: Postmortem redistribution (PMR) is a major challenge in forensic toxicology, particularly when drug concentrations are measured in central blood rather than peripheral blood. Although peripheral blood remains the preferred specimen for postmortem interpretation, it is not always available in routine forensic practice. Methods: This retrospective descriptive study evaluated postmortem central blood drug findings from judicial autopsies analyzed at the Forensic Toxicology Laboratory of the University of Catania between 2017 and 2024. Approximately, 700 forensic toxicological investigations were reviewed. Overall, 52 therapeutic drugs were identified, resulting in a final dataset of 313 individual drug findings. For each finding, central blood concentrations were compared with published reference ranges and cautiously classified as normal/therapeutic, high but non-toxic, toxic, or potentially lethal. Results: Most findings were classified as normal/therapeutic (84.3%), while 28 (8.9%) were high but non-toxic, 15 (4.8%) toxic, and 6 (1.9%) potentially lethal. The distribution remained broadly consistent in sensitivity analyses restricted to drugs represented by more than 3, 5, or 8 findings. In cases with toxic or potentially lethal findings and available paired peripheral blood data, observed C/P ratios were moderate, ranging from 1.07 to 2.15. Conclusions: These findings do not indicate that central blood can replace peripheral blood or that PMR can be disregarded. However, they suggest that central blood drug concentrations are not intrinsically uninterpretable and may retain practical forensic value when interpreted cautiously within the broader medico-legal context. Central blood results should not be used in isolation to diagnose poisoning, estimate the ingested dose, or reconstruct exact ante mortem concentrations.

Forensic Sciences doi: 10.3390/forensicsci6020051

Authors: Everaldo Attard Ilaria Ammendolia Fabrizio Calapai Gioacchino Calapai

Caffeine is the world’s most widely consumed psychoactive compound, yet it presents a growing challenge in forensic toxicology due to the proliferation of highly concentrated energy drinks, dietary supplements, and pure anhydrous powders. This review provides a comprehensive evaluation of the diverse sources of caffeine, complex pharmacokinetics, and critical medico-legal implications. While moderate consumption is socially accepted, fatal intoxications are increasingly reported. The toxic effects of caffeine primarily manifest through a number of mechanisms which range from mild effects to sudden cardiac death. Forensic interpretation is inherently complex due to significant inter-individual variability. Analytical determination relies on advanced techniques, with LC-MS/MS serving as the preferred platform for its high sensitivity and ability to quantify primary metabolites like paraxanthine. A critical challenge in postmortem cases is postmortem redistribution, where central blood levels may be artifactually elevated compared to peripheral sites, necessitating more complex matrix analysis. To standardize the evaluation of the role of caffeine in causing death, a proposed Toxicological Significance Score (TSS) framework is discussed to enhance the transparency and defensibility of expert testimony. Ultimately, accurately determining the manner of death requires a nuanced, case-by-case assessment that integrates toxicological findings with circumstantial evidence and individual medical histories.

Forensic Sciences doi: 10.3390/forensicsci6020050

Authors: Witsarut Upalananda Sawrawit Chairat Sangsom Prapayasatok Sakarat Nalampang Sitthichok Chaichulee

Background/Objectives: Accurate age estimation is a critical task in forensic and legal contexts, where reliable and objective assessment is essential for age-dependent decision-making. This study aims to explore the application of convolutional neural network-based regression models for age estimation from panoramic radiographs, incorporating conformal prediction to generate prediction intervals representing uncertainty. The estimated ages were also evaluated for their application in legal age classification at thresholds of 10, 15, 18, and 20 years. Methods: A total of 1734 panoramic radiographs of individuals aged 8 to 23 years were used in this study. We employed six models, including ResNet-50, EfficientNet-B0, DenseNet-121, MobileNetV3-Small, VGG-16, and ResNeXt-50, to perform regression and provide point estimates of chronological age. Conformal prediction using a split conformal framework with absolute residual scores was implemented to construct statistically valid prediction intervals with finite-sample coverage guarantees. Results: The results revealed that ResNeXt-50 achieved the lowest mean absolute error for point estimates (0.92 years) and produced among the narrowest conformal margins, ranging from 1.71 to 3.72 years across non-coverage probabilities from 0.20 to 0.01. In legal age classification, the ResNeXt-50 model achieved its highest classification accuracy using point estimates, whereas the application of the conformal lower bound improved specificity and positive predictive value. This shift facilitates conservative decision-making, aligning with the forensic minimum-age principle. Conclusions: The proposed framework offers a practical solution for automated age estimation by quantifying uncertainty through prediction intervals, allowing for more informed assessments aligned with forensic protocols for legal age determination.

Forensic Sciences doi: 10.3390/forensicsci6020049

Authors: Raghad Al-Mojahed Nazik Ahmed Rawan Al-Busaidi Adham Al-Rahbi Ali Abduwani Eiman Al-Ajmi Choy Ker Woon Mohamed Al Mushaiqri Srinivasa Rao Sirasanagandla

Background: Accurate sex estimation from skeletal remains is essential in forensic identification, yet population-specific reference data for cervical vertebrae, particularly the first cervical vertebra (C1), are limited. This lack of data compromises the accuracy of sex estimation in forensic contexts where primary skeletal elements such as skull and pelvis are unavailable. Geometric morphometric (GM) analysis offers a robust alternative by quantifying shape variation independent of size, enabling precise assessment of morphological differences that conventional linear measurements may overlook. Objective: This study aimed to investigate the utility of C1 vertebra morphology for sex estimation among Omani adults using two-dimensional geometric morphometrics. Methods: A total of 320 lateral cervical radiographs (160 males, 160 females) were retrospectively collected from Sultan Qaboos University Hospital. Six anatomical landmarks on the atlas were digitized using TPSDig2, and shape analyses were performed using MorphoJ software. Statistical methods included Principal Component Analysis (PCA), Procrustes ANOVA, Canonical Variate Analysis (CVA), and Discriminant Function Analysis (DFA). Intra- and inter-observer reliability was also assessed. Results: While centroid size did not significantly differ between sexes (p = 0.2432), shape analysis revealed statistically significant sexual dimorphism (p = 0.0457). DFA correctly classified 77.19% of individuals by sex, with a cross-validated accuracy of 73.44%. Shape variation was most pronounced in the posterior tubercle and superior facets of the atlas. Conclusions: Two-dimensional geometric morphometric analysis of the first cervical vertebra (C1) reliably captures sexual dimorphism in the Omani population, providing a population-specific reference that enhances the accuracy and objectivity of forensic sex estimation.

Forensic Sciences doi: 10.3390/forensicsci6020048

Authors: Francesco Orsini Stefano Ferracuti Chiara Fabrello Karidia Karaboue Luigi Cipolloni Stefania De Simone

Background/Objectives: Mass suicide events represent complex socio-cultural phenomena requiring multidisciplinary investigation. The 1978 Jonestown event (>900 deaths) remains the most extensively documented modern case. Methods: We conducted a systematic literature review adhering to PRISMA guidelines, searching PubMed, Web of Science, and Scopus databases for peer-reviewed articles published between 1978 and 2026 using the query: ((mass) AND (suicide)) OR (suicide) AND (jonestown). Following screening, 10 papers were selected for inclusion. We integrated these findings with forensic autopsy reports (n = 7), toxicological analyses, discourse analysis of the 45 min audio recording preceding the mass deaths, field investigation data, scholarly monographs, reputable popular science publications, and archival materials from contemporary sources to examine the forensic and criminological aspects of the event. Results: Toxicological analysis detected cyanide in 2/7 autopsied bodies; multiple sedatives (promethazine, chlorpromazine, diphenhydramine, pentobarbital) in gastric tissue indicated recent ingestion. Two gunshot fatalities presented additional complexity. Manner of death was classified as undetermined due to ambiguous circumstances regarding volition versus coercion. Discourse analysis revealed the community’s response was actively debated and constructed through persuasive communication: Reverend Jones strategically framing the situation as hopeless while preserving the appearance of individual agency. Psychosocial analysis identified characteristic patterns: charismatic authoritarian leadership, progressive isolation, apocalyptic ideology, and deindividualization processes. Conclusions: The Jonestown event exemplifies the intersection of forensic pathology challenges in mass casualty investigations and complex group dynamics culminating in collective death. Analysis demonstrates group members actively participated in leadership–followership dynamics, constructing narrative fit between identity and action through discourse. Findings emphasize the importance of immediate toxicological sampling, preservation of forensic evidence, recognition of warning signs in high-control groups, and understanding how narratives of hopelessness and foreclosure of options can be exploited for extremist mobilization.

Forensic Sciences doi: 10.3390/forensicsci6020047

Authors: Silvia Polver Andrea Bobbio Alessandro Angrilli

Background: Stalking is a well-established and studied crime in the forensic field. Nevertheless, research on the psychological aspects of stalking behaviors remains limited due to the lack of specific assessment tools. This two-phase research project aims to propose and validate a new instrument—the Stalking Assessment Questionnaire (SAQ)—to identify the possible latent psychological dimensions characterizing early stalking tendencies. Methods: During the first phase of questionnaire development, a thematic focus group comprising five psychologists generated 55 items. These items underwent preliminary screening with a sample of 85 students to assess clarity and redundancy; this process addressed semantic issues and resulted in a revised 48-item version. In the second validation phase (Study 1), the 48-item SAQ was administered to a sample of 349 male students whose demographic profiles matched those typically associated with potential stalkers; following this, 15 items were retained. In Study 2, the 15-item SAQ was cross-validated on an independent sample of 380 male students. To assess its validity, correlations were analyzed with a battery of personality inventories, including the LSRP, ECR-R, AQ, PESES, SRSS, RelRQ, ASI-S, and BIDR-6. Results: Following item and factor analyses, a model comprising two latent factors—Insistence (SAQ-INS) and Exaggerated Jealousy (SAQ-EXJ)—and 15 indicators emerged. Internal consistency was robust, with a Cronbach’s alpha of 0.87. In Study 2, as hypothesized, SAQ-EXJ correlated positively with anger and hostility (AQ) and with antisocial and impulsive traits (LSRP-F2), although these associations were modest (r = 0.30). Among the significant Pearson’s correlations, three were found to be substantial (r > 0.50): specifically, those between the SAQ and anxious attachment style (ECR-Ranx), relational rumination (RelRQ), and sexism (ASI-S). Conclusions: In conclusion, the final SAQ structure revealed two primary factors—Insistence and Exaggerated Jealousy—that account for stalking as a continuous construct. These factors demonstrated significant associations with several critical personality traits. The final questionnaire comprises 24 items, including nine fillers; consequently, it is efficient to administer and suitable for both research and psycho-educational interventions among adolescents and other potentially vulnerable populations.

Forensic Sciences doi: 10.3390/forensicsci6020046

Authors: Gebremariam Tewelemedhin Gebremariam Charles Karangwa Innocent Nkurunziza David Ishimwe

Background and Forensic Significance: Intimate partner violence (IPV) is a major public health concern that can result in intimate partner homicide (IPH). While most reported cases involve female victims, fatal violence against men, particularly older adults, remains under-recognized. We report an atypical case of IPH involving an elderly male victim who was killed by his female partner. This case highlights the importance of comprehensive medico-legal investigation to accurately reconstruct the mechanism of injury and exclude alternative causes of death. Case Presentation: A 76-year-old man was found dead in his home after his 72-year-old wife reported that she found him unresponsive. He had a history of asthma and chronic alcohol use, was reportedly intoxicated, and had been involved in a longstanding conflict with his spouse. Autopsy revealed multiple patterned abrasions and contusions with extensive intramuscular hemorrhage involving the extremities. Internal examination demonstrated generalized visceral pallor. Fatal acute asthma and alcohol intoxication were excluded. Overall, the cause of death was determined to be hemorrhagic shock resulting from repeated blunt force trauma, and the manner of death was classified as homicide. Conclusions: This case demonstrates that a sustained blunt force assault can lead to fatal intramuscular blood pooling. Furthermore, it illustrates the rare demographic and mechanism patterns of IPH, underscoring the importance of comprehensive medico-legal evaluation and awareness of such occurrences.

Forensic Sciences doi: 10.3390/forensicsci6020045

Authors: Yara Vieira Lemos Ricardo Moreira Araújo Felippe Bevilacqua Prado Alexandre Rodrigues Freire Ana Cláudia Rossi

Background/Objectives: The frontal sinus exhibits individual morphological variability that may support human identification. Its development progresses through childhood and adolescence and stabilizes in early adulthood, with age-related changes potentially affecting radiological comparisons. This study presents a forensic case report and discusses it in light of the literature on frontal sinus development and forensic identification. Methods: A comparative radiological analysis was conducted using images obtained at two distinct stages of biological maturation (14 and 21 years of age). Manual delineation combined with semi-automated computational analysis was applied to assess morphological features of the frontal sinus, including contour configuration, lobulation, and dimensional parameters. Results: The intra vitam record was obtained at 14 years of age, during an active developmental phase, and the post mortem examination was obtained at 21 years, corresponding to early adulthood. Comparative analysis revealed significant morphological differences, including increased lobulation, contour complexity, and sinus expansion. These changes limited the reliability of frontal sinus morphology for identification in this case. Friction ridge examination independently established positive identification. Conclusions: This study highlights the limitations of frontal sinus analysis when applied across periods of active development and underscores the risk of misinterpretation if age-related changes are not adequately considered.

Forensic Sciences doi: 10.3390/forensicsci6020044

Authors: Yangseung Jeong Lee Meadows Jantz Yochun Jung

Background: Accurate estimation of the pre-colonization interval (pre-CI), defined as the period between body deposition and initial insect oviposition, remains a challenge in postmortem interval estimation in forensic entomology. This study quantified the effects of environmental and contextual variables on blowfly oviposition timing using human cadavers. Methods: Daily photographic records from 203 donated cadavers placed at the University of Tennessee Anthropology Research Facility (March 2011–July 2014) were used to document the first observed oviposition of blowflies. Multivariable general linear models evaluated the effects of monthly temperature, black plastic coverage, and pre-placement soft tissue damage on calendar days to oviposition (IOday) and accumulated degree days (IOADD). Results: Temperature was the dominant predictor, explaining substantial variation in pre-CI. Black plastic coverage significantly delayed oviposition and exhibited a temperature-dependent effect. Although soft tissue damage was associated with earlier oviposition in univariable analyses, it did not remain significant in multivariable models. Cold conditions were associated with prolonged and highly variable pre-CI. Conclusions: These findings demonstrate that temperature was the strongest predictor in this observational dataset, while contextual factors such as physical barriers modify colonization patterns. Together, they highlight the need to incorporate environmental and contextual variables into PMI estimation models and support more defensible interpretations of entomological evidence, particularly in cases involving delayed colonization or restricted insect access.

Forensic Sciences doi: 10.3390/forensicsci6020043

Authors: Eva Ronner Nicholas Márquez-Grant Amaya Gorostiza Jorge Moreno Andrés Alfonso M. Villalta Luna Julián López García María García Alonso Isabel Angulo Bujanda María García Velasco María Benito Sánchez

Introduction: During an excavation in 2022 of a mass grave within a cemetery in Castilla-La Mancha containing the remains of executed victims of the Spanish Civil War and Francoist dictatorship, the scientific team discovered a medallion with a name and a date inscribed on it. The medallion was associated with an individual recovered from a grave containing 12 other individuals in 5 separate stratigraphic layers. This paper aims to outline the identification process of the deceased from the skeletal remains and associated medallion. Methods: This process included archival research, gathering witness testimonies, archaeological evidence, osteological examination and DNA analysis. Results: After restoration, the medallion revealed the words “VIVA LA FIJL. VIVA LA FAI.” on one face and “MUERO POR LA LIBERTAD.” on the other. The human skeletal remains associated with the medallion showed male morphological characteristics and were estimated to be between 21 and 30 years old at the time of death, with a height between 1.64 and 1.67 m. There was skeletal evidence of ballistic trauma on the right mandible and the right scapula. Genetic analysis confirmed an mtDNA match with his maternal niece. Discussion/Conclusions: This study demonstrates the importance of a multidisciplinary approach to the identification of human remains from the Spanish Civil War and the Dictatorship which followed, and how the objects found within a mass grave can be useful in aiding a positive identification.

Forensic Sciences doi: 10.3390/forensicsci6020042

Authors: Simona C. Ionescu Sebastian R. Cozma Irina S. Manoilescu Eugen C. Ionescu Alexandra C. Neagu

Post-traumatic auditory and vestibular complaints are frequent after traumatic brain injury (TBI) and temporal bone trauma. They create particular difficulty in medico-legal practice because the evaluator must distinguish diagnosis, functional impact, plausibility of traumatic causation, and the credibility of reported deficits and/or symptoms. This manuscript is a narrative expert review, not a systematic review or a validated forensic prediction rule. It aims to synthesize clinically relevant evidence and propose a practical cross-check framework for structured audio-vestibular assessment in post-traumatic and medico-legal contexts. Pure-tone audiometry remains the functional entry point, but it should be interpreted in conjunction with speech audiometry, tympanometry, acoustic reflexes, transient-evoked and distortion-product otoacoustic emissions, auditory brainstem responses, and auditory steady-state responses. Vestibular evaluation should combine videonystagmography, video head impulse testing, cervical and ocular vestibular evoked myogenic potentials, and computerized dynamic posturography, recognizing that each method interrogates different physiological domains and frequencies. Particular emphasis is placed on the separation between clinical diagnosis, physiological localization, functional impairment, and medico-legal attribution. The article also discusses safeguards against false-positive attribution of malingering, the time course after TBI, inter-rater variability, and the role of specialist expertise in medico-legal reporting. The proposed framework does not eliminate uncertainty; rather, it is intended to make expert reasoning transparent, cautious, internally consistent, and defensible.

Forensic Sciences doi: 10.3390/forensicsci6020041

Authors: Muhammet Can Cihangir Işık Meksel Cengiz

Background/Objective: Forensic document examination (FDE) traditionally relies on subjective expert opinion. This preliminary study was designed to develop and validate a hybrid deep learning model (ResNet-50 + bidirectional long short-term memory [BiLSTM]) for quantitative handwriting and signature feature analysis, and to compare its performance, under standardized experimental conditions, with that of three certified forensic document examiners. Methods: Handwriting and signature samples were collected from 225 individuals in a standardized setting. Fifteen quantitative handwriting features were extracted, the dataset was split into training (70%, n = 158) and testing (30%, n = 67) subsets using stratified random sampling, and ground truth for analytic categories was defined by majority consensus among the three examiners (with adjudicated review for disagreements). A hybrid architecture combining a ResNet-50 backbone and a bidirectional LSTM encoder was used. Results: The model demonstrated 93.4% accuracy, an F1-score of 0.926, and an AUC-ROC of 0.968 on the held-out test set. Under our task-specific experimental conditions, the model performed better than examiners on slant analysis (96.8% vs. 93.2%, p = 0.002), pressure profiling (94.1% vs. 91.7%, p = 0.019), and age estimation (87.4% vs. 82.1%, p = 0.011); examiners performed better on forgery detection (95.8% vs. 91.2%, p = 0.008) and signature verification (96.1% vs. 92.3%, p < 0.012). Mean processing time was reduced by 99.6% (0.8 s vs. 197 s per case). Conclusions: Within the limits of this preliminary single-centre study, the system showed performance comparable to certified examiners on several quantitative tasks and complementary strengths overall, supporting its feasibility as an adjunctive tool in a hybrid human–AI workflow. Broader, multi-centre validation and explainability work are required before any forensic deployment can be considered.

Forensic Sciences doi: 10.3390/forensicsci6020040

Authors: Ana Leonardo Rita Files Gonçalo da Graça Pereira Isabel Pires

Background/Objectives: Dog tethering remains a common containment practice in several regions despite increasing concerns about its implications for animal welfare. This study aimed to assess veterinary professionals’ perceptions in Portugal about the impact of dog tethering on canine health, behavior, and welfare, as well as their views on its regulation. Methods: An anonymous, structured questionnaire was distributed online to veterinary professionals to collect data on clinical exposure to chained dogs, observed health and behavioral outcomes, and attitudes toward criminalizing the practice. Results: A total of 331 veterinarians participated, of whom 71.9% reported encountering chained dogs during their professional activities, particularly in rural areas. Most respondents perceived the tethering conditions as inadequate, particularly regarding freedom of movement and the fulfillment of behavioral needs. Frequently reported consequences included alopecia at the restraint site, poor muscle condition, physical injuries, fear-related behaviors, reduced socialization, and increased aggression. Preventing escape was identified as the primary perceived motivation for tethering. The majority of veterinarians supported some degree of criminalization, particularly when welfare impairment was evident. Conclusions: These findings reflect veterinary professionals’ perceptions of the welfare risks associated with prolonged tethering and underscore the need for integrated strategies combining regulation, education, and accessible, humane containment alternatives.

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.