Formalin pigment is the most important artifact found in postmortem histological analyses. Although it has no forensic value, formalin pigment, when abundant or not well-known to the pathologist, can morphologically resemble, mimic or misinterpret physiological or exogenous pigments of forensic interest, making their identification challenging and leading to diagnostic difficulties. To investigate these aspects, a retrospective study was conducted on 35 autopsied bodies in various states of preservation in which postmortem histological analyses had revealed the presence of brown/blackish precipitates requiring a differential diagnosis with formalin pigment artifacts. In 29 cases (83%) it could be proven that the precipitates were not due to formalin artifacts. In some cases, however, they were of minor forensic significance, so that others could be categorised as highly forensically significant. These were mainly haemosiderin, lead from gunshot residue (GSR), granular precipitates as a result of acute poisoning with various substances and malaria pigment. It should therefore be noted that precipitates that can be superficially assigned to formalin artifacts can conceal a multitude of pigments of different types, aetiologies and thus forensic significance, which also depend on the circumstantial and investigative context of the individual cases. For this very reason, it is of central importance for forensic pathologists to constantly question whether it may be useful to further investigate the diagnosis of suspected artifactual formalin pigment precipitates in autopsy specimens.

Cardiac metastases from malignant melanoma are rare and often remain undiagnosed until autopsy due to their nonspecific clinical presentation. The case report concerns a woman in her 40 s, with medical history of coeliac disease who presented with a triad of cholestatic jaundice, weight loss and generalised weakness. Acute cardiac symptoms occurred during the patient's hospitalisation, with dyspnoea and palpitations preceding refractory ventricular arrhythmias and sudden death. Post mortem examination showed extensive metastatic melanoma that included the lungs, liver, spleen, lymph nodes and notably the myocardium and the endocardium. Importantly, no cutaneous or mucosal primary site was identified, classifying this case as melanoma of unknown primary (MUP). The histological and immunohistochemical analysis confirmed the diagnosis. This case report summarises the range of diagnostic issues associated with cardiac metastasis from melanoma, where the clinical presentation can mimic other cardiac pathologies and cause undiagnosed fatal arrhythmias. The findings emphasise the significance of meticulous autopsy and histological examination in elucidating unrecognized cardiac involvement in patients with metastatic melanoma, particularly in MUP.

Body temperature is an important indicator for forensic scientists to estimate the early post-mortem interval (PMI). Traditionally, forensic practitioners have mostly used methods such as measuring the internal temperatures of the body, including rectal temperature and liver temperature, to infer the PMI. However, these methods rely on the experience of the operator, suffer from manual measurement errors, and cannot fully consider the interference of environmental factors, resulting in unstable accuracy of the inference results. Infrared thermography technology can use optoelectronic technology to dynamically and monitor the surface temperature of objects and ambient temperature. It has the advantages of non-contact, convenience, and high efficiency. This project utilized thermography technology to monitor the surface temperature of SD rat corpses and the real-time ambient temperature under five average environment temperature conditions of 25 °C, 20 °C, 15 °C, 10 °C, and 5 °C, aiming to explore the correlations among the changes in body surface temperature, ambient temperature changes, and PMI. By comparing the changes in body surface temperature of rabbits, SD rats, and C57 mice, the project investigated the relationship between body weight and the rate of body surface temperature decrease. Then, multivariate correlation analysis, multiple regression analysis, and machine learning analysis methods were employed to construct and evolve models for PMI estimation. Finally, the accuracy of the model was verified using the single-blind method. The results showed that using infrared thermography to detect body surface temperature and ambient temperature can more conveniently and accurately infer the PMI, demonstrating high application and transformation value.

Botulinum toxin type A is widely used to block acetylcholine release in the treatment of chronic sialorrhea, muscle spasticity, and dystonia. We aim to develop a user-friendly method for detecting cases of medical botulinum toxin poisoning.

Impacted mandibular third molars (IMTM) are often excluded from dental age estimation due to limited accessibility and morphological variability. This study aimed to develop an age estimation model based on the apical surface area (ASA) of IMTM roots using cone-beam computed tomography (CBCT). A total of 446 CBCT images of Malaysians aged 15-25 years (243 females, 203 males) from Malay and Chinese ethnicities were retrospectively analyzed. ASA measurements were obtained using Mimics and 3-matic software (Materialise NV, Belgium). Impaction levels were classified via a modified Pell and Gregory system, and impaction types using Quek's modification of Winter's classification. IMTM angulation, root number, and orientation across three planes were assessed. ASA means were compared by sex and ethnicity using t-tests, and ANOVA was used to evaluate differences across impaction levels, types, and root numbers. General linear regression was applied to identify significant predictors of chronological age and to develop the model, which was validated using an additional 123 CBCT scans. ASA showed a strong inverse correlation with age (r = -0.73). Significant sex-based differences in ASA were observed (male: p = 0.02; female: p = 0.03), with no significant differences between ethnicities. ASA, sex, number of roots, and impaction type were significant predictors of age. The model achieved an R of 0.607 and mean absolute error ranging from 0.85 to 4.87 years across age groups. This study underscores the use of IMTM from CBCT data as a feasible method for legal age estimation in the Malaysian population, particularly in distinguishing juveniles from adults.

Female genital mutilation (FGM) refers to socio-cultural practices involving the partial or total removal of a woman's external genitalia for non-medical purposes. These procedures are associated with numerous short- and long-term physical and psychological complications. While such complications are relatively well documented in adults, pediatric cases remain insufficiently reported. Forensic physicians are often the first - and sometimes the only - healthcare providers to detect and document these complications, particularly in asylum contexts.

Diatoms inhaled with a drowning medium are thought to reach closed organs, including the kidney and liver, through circulation; however, this is uncertain. Thus, to assess this doubt, we evaluated the relationship between resuscitation or decomposition and diatom concentration in closed organs (kidney and liver) (Cc).

This study investigates the postmortem tissue distribution of morphine, its main metabolites, and levetiracetam in a terminal cancer patient. A validated liquid chromatography-tandem mass spectroscopy method was developed and applied to blood and tissue samples collected during a forensic autopsy. Samples were processed using protein precipitation with acetonitrile and lipid removal cartridges. The analysis revealed peripheral blood concentrations of 0.220, 1.19, and 16.6 μg/mL for morphine, morphine-3-glucuronide/morphine-6-glucuronide, and levetiracetam, respectively. The highest concentrations of each drug were found in the urine. The concentration of free morphine in the blood was elevated, but higher levels have been reported in terminal cancer patients undergoing long periods of pain management with morphine. Levetiracetam blood levels were in the therapeutic range. The results align with existing literature on morphine's postmortem redistribution and provide new insights into the tissue distribution of levetiracetam. These data can serve as a reference for future studies and contribute to the development of guidelines for the use of these medications in end-of-life care. The study concludes that while the drug concentrations were not high enough to solely attribute the cause of death to intoxication, the comprehensive tissue distribution analysis offers valuable information for forensic and clinical applications. To our knowledge, this is the first report to document the simultaneous analysis and tissue distribution of morphine and levetiracetam in postmortem samples.

A one-month-old girl in good health being carried by her mother in a baby carrier suddenly went into cardiac arrest. She was rushed to the hospital, but was pronounced dead. A forensic autopsy was performed. On the heart, which weighed 63.6 g, a single large, well-defined, round, white tumor was observed in the left ventricular wall. Tumor cells with spindle-shaped small nuclei proliferated along with collagen fiber proliferation. The diagnosis was cardiac fibroma. The tumor had compressed the left bundle branch, causing fatal arrhythmia and death. This case report describes sudden infant death caused by a primary cardiac fibroma, diagnosed postmortem.

DNA analysis in forensic science involves short tandem repeat analysis using genomic DNA, however, it is limited by low DNA concentration. Then, mitochondrial DNA (mtDNA) can be used as an alternative. In this study, to analyze mtDNA control region, we conducted an analysis using Sanger sequencing and two NGS methods: ForenSeq mtDNA Control Region Kit and Precision ID mtDNA Control Region Panel. This study is a comparative analysis of two commercially available NGS kits for mtDNA control region analysis. MtDNA was extracted from 75 Korean samples, and sequenced using Sanger sequencing, ForenSeq mtDNA Control Region Kit, or Precision ID mtDNA Control Region Panel. Although most mtDNA control region showed identical results between Sanger sequencing and NGS, differences in particular regions were observed in 43 of 75 samples using Sanger sequencing, ForenSeq mtDNA Control Region Kit, and Precision ID mtDNA Control Region Panel. These sites were poly-C sites of 16184-16193 in HV1, 303-315 in HV2, and 568-573 in HV3 that have been observed in InDel or substitution variant frequently. Additionally, heteroplasmy that was not observed in Sanger sequencing was detected in 6 samples in the NGS analysis. Therefore, mtDNA analysis using NGS is a convenient method for analyzing a large number of samples at once and is advantageous for detecting heteroplasmy, however, in length-based variation region, as bias can occur due to potential sequencing or alignment artifacts, caution should be taken in the interpretation of repetitive regions.

The rupture of coronary artery plaque is a pathological mechanism leading to acute coronary syndrome, typically associated with severe atherosclerosis and high hemodynamic stress, while cases induced by blunt chest trauma (BCT) are exceedingly rare. This report presents an unusual case of a young male who died shortly after an informal basketball game. Postmortem examination revealed mild atherosclerotic plaque with a lacerated fibrous cap, along with complete thrombotic occlusion in the proximal left anterior descending (LAD) artery. Some areas of the thrombus were in direct contact with the lipid and necrotic components of the plaque. Witness testimony indicated the presence of BCT. Moreover, focal epicardial hemorrhage around the LAD artery, along with microscopically identified subcutaneous hemorrhage in the chest wall skin further supported the occurrence of BCT. Ultimately, the cause of death was determined to be BCT-related plaque rupture and the subsequent thrombus formation in the LAD artery. Given the rarity of this phenomenon, the association between BCT and plaque rupture requires careful clarification and may lead to forensic disputes. This report aims to remind forensic pathologists to consider the possibility of plaque rupture following BCT when faced with similar cases.

Accurate detection and quantification of human nuclear DNA (nDNA), named the human accelerated region 1 (HAR1) assay, was developed to simultaneously prove the human origin of forensic biological samples and optimize nDNA input for PCR in human DNA profiling. The HAR1 assay, a TaqMan probe-based real-time PCR assay targeting HAR1 in nDNA, was exclusively specific to human DNA samples with no cross-reactivity across 23 vertebrate species (mammals including non-human primates, birds, reptiles, amphibians, and fishes). This assay is capable of proving the human origin of the samples from as little as the DNA content of a single somatic cell. Moreover, accurate quantification of human nDNA was achieved down to the minimum input concentration (33 pg/μL) recommended by commercial DNA profiling kits. Furthermore, it was confirmed that the presence of non-human DNA mixed with human DNA did not interfere with the accuracy of human nDNA quantification. Therefore, the HAR1 assay represents a valuable and practical tool for improving both the accuracy and efficiency of human DNA profiling, particularly in forensic casework where sample quantity and quality may be limited and the presence of non-human DNA can complicate analysis.

Venous air embolism (VAE) secondary to basilar skull fractures is rare but potentially fatal. While VAE is a known complication in central venous catheter management, traumatic VAE-particularly in forensic contexts-remains underreported. In forensic medicine, few case reports have detailed the use of postmortem computed tomography (PMCT) to diagnose VAE.

Liposuction remains one of the most commonly performed cosmetic surgical procedures worldwide, yet fatal complications continue to occur despite advances in surgical techniques. This study aims to conduct forensic pathological analysis of liposuction-related deaths to identify specific complications, underlying causes, and patterns of fatal outcomes.

Tuberculous myocarditis is a rare manifestation of disseminated tuberculosis that is often misdiagnosed. It may present with non-specific symptoms, or alternatively remain clinically silent, making diagnosis particularly challenging. We present in this paper a case report of a 49-year-old man with a medical history of diabetes, untreated hypertension, and dyslipidemia was found dead in his home. There was no specific symptom before death. A medico-legal autopsy was requested to determine the cause of death. At the internal examination, there were bilateral yellow-citrine pleural effusion, pulmonary edema with congestion, whitish areas in the left ventricular myocardium. Histopathological analysis showed granulomatous myocarditis associated with interstitial granulomatous pneumonitis suggestive of miliary tuberculosis. The cause of death was attributed to acute heart failure related to undiagnosed tuberculous myocarditis.

Clarifying the cause of death of heat stroke (HS) is critical for judicial handling of related cases. Yet, postmortem diagnosis of HS remains challenging owing to the lack of specific morphology and biomarkers. Liver damage frequently acts as a direct cause in HS. This study aimed to explore the postmortem diagnosis of HS based on the protein biomarkers in liver by machine learning methods.

Ingested alcohol (ethanol) is oxidized in the liver to acetic acid, which is then transferred to other organs where it is completely oxidized to carbon dioxide and water. However, to the best of our knowledge, no overall equation for this process has yet been described in the forensic literature, despite its physiological relevance. Here we present such an overall equation, derived by summing the reactions involved in the complete oxidation of ethanol: oxidation of ethanol to acetic acid by alcohol and aldehyde dehydrogenases, conversion to acetyl-coenzyme A, oxidation via the citric acid cycle, and utilization of the resulting reduced form of nicotinamide adenine dinucleotide and ubiquinol to fully reduce molecular oxygen. The overall equation is expressed as a pair of equations because inorganic phosphate takes two forms, HPO and HPO, with a pKa of approximately 6.8. The pair of equations indicates that no H is generated if inorganic phosphate is present as HPO, whereas one H is formed if inorganic phosphate is present as HPO. The actual ratio of [HPO]/[HPO] is approximately 1, 1.6, and 2.5 at pH 6.8, 7.0, and 7.2, respectively, indicating that complete oxidation of one ethanol molecule forms approximately 0.6 (0.5-0.7) H. Evidence of H formation contradicts the general notion that complete oxidation neither generates nor consumes H. The implications of H generation merit clarification, particularly given the prevalence of metabolic acidosis in heavy drinkers.

We developed a novel next-generation sequencing-based genotyping panel for 200 single nucleotide polymorphism (SNP) loci, and evaluated the genotyping performance when using small amounts of DNA. We analyzed 1144 previously characterized autosomal SNPs, and selected 200 informative SNPs, based on the allele coverage ratio (ACR) and amplicon size, to minimize inter-locus depth of coverage imbalances. Analysis of the 200 SNPs was performed using 1 ng of DNA from 66 Japanese individuals. The average ACR of most of the SNP loci ranged from 0.85 to 0.95. We also detected 200-SNP genotypes using diluted DNA samples (500, 250, 125, 60, 30, and 15 pg/reaction), and found high ACRs and concordance from less than 100 pg of DNA: the concordances were as high as 95.7 % and 88.0 %, respectively, when 60 and 30 pg of DNA were used. Our method also showed higher inhibitory tolerance than other methods when humic acid was added to the initial amplification reaction for SNP genotyping. When genotyping in the presence of 10 ng/µL humic acid, no SNP profiles were detected, but profiles with more than 99 % concordance were recovered on addition of bovine serum albumin. The genotyping performance was also evaluated using small groups of cells (10, 5, 4, 3, and 2 cells/reaction) isolated by micromanipulation with micro-tweezers. High concordance was identified on analysis of three cells (96.1 %). The use of isolated cells allowed accurate determination of the sensitivity, indicating that our method is highly sensitive and can provide informative SNP profiles for individual identification.

Bland-Garland-White syndrome, or an anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA), is a rare congenital coronary anomaly that can cause myocardial ischemia, congestive heart failure, and sudden death. In this study, we aimed to present a case of sudden infant death caused by ALCAPA. Postmortem computed tomography (PMCT) showed a collapsed left lung, enlarged heart volume, and severe calcification of the left ventricular papillary muscles and posterior left ventricular wall. Macroscopic examination during the autopsy revealed marked cardiac hypertrophy and an anomalous left coronary artery originating from the pulmonary artery. Furthermore, microscopic examination showed significant myocardial fibrosis with severe left ventricular calcification. Histological findings revealed features of acute-on-chronic myocardial ischemia, including myocytolysis around fibrotic and calcified areas, contraction-band necrosis, and a characteristic wavy cardiomyocyte pattern. In this case, we suggest that localized myocardial calcification detected by PMCT, particularly following three-dimensional visualization, may be useful for the postmortem assessment of ALCAPA and other causes of chronic myocardial ischemia in infants.