A calcification subtraction method for postmortem coronary computed tomography angiography.

Although coronary computed tomography (CT) angiography is a useful tool for evaluating coronary artery lesions both ante- and postmortem, accurate evaluation of the lumen is difficult when highly calcified lesions are present, owing to overestimation of stenosis caused by blooming and partial volume artifacts. In clinical practice, to overcome this diagnostic problem, a subtraction method has been devised to remove calcification by subtracting the precontrast image from the contrast image. In this report, we describe a calcification subtraction method using image analysis software for postmortem coronary CT angiography. This method was devised based on preliminary experimental results showing that the most accurate subtraction was achieved using images reconstructed with a narrower field of view and bone kernel, resulting in higher spatial resolution. This subtraction method allowed evaluation of lumen patency and the degree of stenosis on contrast-enhanced images in a verification using actual specimens where evaluation of the lumen had been difficult because of high calcification. The results were morphologically similar to the macroscopic findings. This method allows more rapid and reliable lesion retrieval and is expected to be useful for postmortem coronary angiography in forensic practice.

Sex estimation from skull measurements of a contemporary Japanese population using three-dimensional computed tomography images.

In this study, we assessed the sexual dimorphism of the contemporary Japanese skull and established sex discriminant function equations based on cranial measurements using three-dimensional (3D) computed tomography (CT) images. The CT images of 263 corpses (142 males, 121 females) that underwent postmortem CT scanning and subsequent forensic autopsy were evaluated. Twenty-one cranial measurements were obtained from 3D CT reconstructed images, which extracted only bone data. We performed descriptive statistics and discriminant function analyses for the measurements. Nineteen measurements were significantly larger in males, suggesting sexual dimorphism of the Japanese skulls. Univariate discriminant function analyses using these measurements showed a sex classification accuracy of 57.8-88.2%, and bizygomatic breadth provided the highest correct prediction rate. Multivariate discriminant function analyses offered the most accurate model using seven variables with an estimation rate of 93.9%. Our results suggest that cranial measurements based on 3D CT images may help in the sex estimation of unidentified bodies in a contemporary Japanese population.

Age estimation by evaluating median palatine suture closure using postmortem CT.

This study sought to develop an age-estimation formula to evaluate the extent of median palatine suture (MP) closure using postmortem computed tomographic (PMCT) images. The PMCT images of 634 Japanese subjects (mean age, 54.5 years; standard deviation [SD], 23.2 years) with known age and sex were examined. The degree of suture closure of the MP, anterior median palatine suture (AMP), and posterior median palatine suture (PMP) was measured and scored (suture closure score, SCS), and a single linear regression analysis was conducted with age at death. On the analysis, SCS of MP, AMP, and PMP showed a significant correlation with age (p < 0.001). The correlation coefficient of MP was higher (0.760, male; 0.803, female; and 0.779, total) than that of AMP (0.726, male; 0.745, female; and 0.735, total) or PMP (0.457, male; 0.630, female; and 0.549, total). The regression formula and standard error of estimation (SEE) of MP were calculated as Age = 100.95 × SCS + 20.51 (SEE 14.87 years) for male subjects, Age = 91.93 × SCS + 26.65 (SEE 14.12 years) for female subjects, and Age = 95.17 × SCS + 24.09 (SEE 14.59 years) for the total, respectively. In addition, another 50 Japanese subjects were randomly selected to validate the age-estimation formula. In this validation, the actual age of 36 subjects (72%) was within the estimated age ± SEE. This study showed that the age estimation formula using PMCT images of MPs was potentially useful for estimating the age of unidentified corpses.

Stature estimation by semi-automatic measurements of 3D CT images of the femur.

Stature estimation is one of the most basic and important methods of personal identification. The long bones of the limbs provide the most accurate stature estimation, with the femur being one of the most useful. In all the previously reported methods of stature estimation using computed tomography (CT) images of the femur, laborious manual measurement was necessary. A semi-automatic bone measuring method can simplify this process, so we firstly reported a stature estimation process using semi-automatic bone measurement software equipped with artificial intelligence. Multiple measurements of femurs of adult Japanese cadavers were performed using automatic three-dimensional reconstructed CT images of femurs. After manually setting four points on the femur, an automatic measurement was acquired. The relationships between stature and five femoral measurements, with acceptable intraobserver and interobserver errors, were analyzed with single regression analysis using the standard error of the estimate (SEE) and the coefficient of determination (R2). The maximum length of the femur (MLF) provided the lowest SEE and the highest R2; the SEE and R2 in all cadavers, males and females, respectively, were 3.913 cm (R2 = 0.842), 3.664 cm (R2 = 0.705), and 3.456 cm (R2 = 0.686) for MLF on the right femur, and 3.837 cm (R2 = 0.848), 3.667 cm (R2 = 0.705), and 3.384 cm (R2 = 0.699) for MLF on the left femur. These results were non-inferior to those of previous reports regarding stature estimation using the MLF. Stature estimation with this simple and time-saving method would be useful in forensic medical practice.

Postmortem magnetic resonance imaging revealed bilateral globi pallidi lesions in a death associated with prolonged carbon monoxide poisoning: a case report.

A man and a woman were found dead in the same car with a burned coal briquette. The cause of death of the woman was assigned to acute carbon monoxide (CO) poisoning without difficulty based on typical findings associated with this condition, including elevation of carboxyhaemoglobin (COHb). However, the man had an unremarkable elevation of COHb and a higher rectal temperature compared to that of the woman. Postmortem computed tomography (PMCT) revealed ambiguous low-density areas in the bilateral globi pallidi. Further analysis by postmortem magnetic resonance (PMMR) imaging showed these lesions more clearly; the lesions appeared as marked high signal intensity areas on both the T2-weighted images and the fluid-attenuated inversion recovery sequences. A subsequent autopsy revealed signs of pneumonia, dehydration, starvation, and hypothermia, suggesting that the man died from prolonged CO poisoning. Both globi pallidi contained grossly ambiguous lesions, and a detailed neuropathologic investigation revealed these lesions to be coagulative necrotic areas; this finding was compatible with a diagnosis of prolonged CO poisoning. This case report shows that postmortem imaging, especially PMMR, is useful for detecting necrotic lesions associated with prolonged CO poisoning. This report further exemplifies the utility of PMMR for detecting brain lesions, which may be difficult to detect by macroscopic analysis.

Sudden death due to tension pneumothorax associated with pulmonary tuberculosis.

A man in his seventies who lived alone was found dead in his home. Postmortem computed tomography (CT) performed prior to autopsy showed right-sided tension pneumothorax. Autopsy revealed an active pulmonary tuberculosis (TB) lesion. Macroscopic and histopathological findings showed pleural infiltration by TB lesions, suggesting that tension pneumothorax developed in association with TB infection. Routine postmortem CT performed prior to autopsy is useful in screening for TB because the presence of TB lesions can be confirmed from characteristic pulmonary findings. However, it may be difficult to identify tuberculous pulmonary lesions on CT if pneumothorax occurs and the lung collapses completely as in this case. Thus, forensic pathologists and radiologists should be cognizant of this rare complication of TB.

Postmortem CT and MRI findings of massive fat embolism.

OBJECTIVE: To elucidate postmortem computed tomography (PMCT) and postmortem magnetic resonance (PMMR) imaging findings suggesting massive fat embolism. MATERIALS AND METHODS: Consecutive forensic cases with PMCT and PMMR scans of subjects prior to autopsy were assessed. For PMCT, 16- or 64-row multidetector CT scans were used; for PMMR, a 1.5 T system was used. MRI sequences of the chest area included T2- and T1-weighted fast spin-echo imaging, T2*-weighted imaging, T1-weighted 3-dimensional gradient-echo imaging with or without a fat-suppression pulse, short tau inversion recovery, and in-phase/opposed-phase imaging. At autopsy, forensic pathologists checked for pulmonary fat embolism with fat staining; Falzi's grading system was used for classification. RESULTS: Of 31 subjects, four were excluded because fat staining for histopathological examination of the lung tissue could not be performed. In three of the remaining 27 subjects, histology revealed massive fat embolism (Falzi grade III) and the cause of death was considered to be associated with fat embolism. CT detected a "fat-fluid level" in the right heart or intraluminal fat in the pulmonary arterial branches in two subjects. MRI detected these findings more clearly in both subjects. In one subject, CT and MRI were both negative. There were no positive findings in the 24 subjects that were fat embolism-negative by histology. DISCUSSION AND CONCLUSION: In some subjects, a massive fat embolism can be suggested by postmortem imaging with a "fat-fluid level" in the right heart or intraluminal fat in the pulmonary arterial branches. PMMR potentially suggests fat embolism more clearly than PMCT.

Detection of diatoms in a case of mud aspiration at a coastal area.

The source of diatoms detected in non-drowned bodies have been attributed to postmortem invasion, contamination during autopsy or diatom testing, or "natural load." However, sand aspiration has not been reported as a source. Herein, we report an autopsy case in which diatoms were detected in a non-drowned human who had aspirated mud. A man in his 60 s was found dead at a harbor park in Japan. His whole body was covered with sand, including his face. A situational investigation suggested that he may have entered the sea. Autopsy revealed intratracheal mud, with no obvious findings indicating drowning, suggesting that he died from mud aspiration probably due to hypothermia or non-lethal ethanol intoxication. In the diatom test, 10-100 diatoms/g were detected in bilateral lung samples, which were similar to those found in the intraoral and intratracheal mud and the sand samples from around the discovery site and not similar to those found in the seawater samples. The diatoms in the stomach content exhibited an intermediate trend between those found in the sand and seawater. Therefore, careful qualitative and quantitative analyses are required to differentiate between true drowning and false-positives in non-drowning cases to determine the cause of death.

Lethal hemopericardium caused by infection of mitral annular calcification: An autopsy case report.

Mitral annular calcification (MAC) is a chronic and degenerative condition involving calcification of the mitral annulus. MAC is a risk factor for coronary artery disease, cardiovascular events, stroke, and cardiovascular death. However, patients with MAC are often asymptomatic. Herein, we present the first case of cardiac tamponade due to infection of MAC in forensic pathology. An 80-year-old woman was found in cardiopulmonary arrest shortly after experiencing fatigue. She was transferred to a hospital, and despite chest compression and ventilation, she was pronounced dead due to no response. Postmortem computed tomography, autopsy, and histological examination showed MAC, abscess formation involving Gram-positive cocci on the MAC, and fistulation of the abscess into the intracardial pericardial cavities, resulting in a massive lethal hemopericardium.

Mandibular torus thickness associated with age: Postmortem computed tomographic analysis.

Age estimation is an essential step in identifying human corpses. Several mandibular landmarks have been highlighted as skeletal sites for age estimation since aging causes morphological changes. Reports suggest that mandibular torus size may be associated with aging; however, thorough investigation has not been performed owing to the difficulty in measuring it. Therefore, this study aimed to determine the association between age and mandibular torus thickness using postmortem computed tomography data from Japanese corpses. This study included 2,792 corpses with mean (standard deviation) age of 58.0 (22.4) years (range, 0-101 years) and 67.6 % males. Further, 2,662 (95.3 %), 14 (0.5 %), 59 (2.1 %), and 57 (2.0 %) corpses were in the permanent, mixed, primary, and predental dentition periods, respectively. Multivariable analysis was performed to quantify the impact of age on mandibular torus thickness, adjusting for sex, height, weight, and occlusal contact status. The model also included an interaction term between age and occlusal status because of the potential effect modification by occlusion. Results of the multivariable regression analysis showed that mandibular torus thickness increased with age (the regression coefficients (95 % confidence interval) were 0.6 (0.2-1.0), 0.7 (0.3-1.0), 1.0 (0.6-1.4), 1.3 (0.9-1.7), 1.3 (0.8-1.8), and 1.1 (0.4-1.7) for age groups 30-39, 40-49, 50-59, 60-69, 70-79, and 80-89 years, respectively), especially in males with occlusal contact. A significant association between mandibular torus thickness and age, modified by occlusal status and sex, was identified. Therefore, data regarding the thickness of the mandibular torus and occlusal status may be useful for age estimation in human corpses.

Assessment of differences in mechanical properties and thickness of temporal and occipital bones and correlation with age in Japanese forensic samples.

The mechanical properties and thickness of adult temporal and occipital bones were examined in modern Japanese forensic samples. Cranial bones were obtained from 293 Japanese corpses (179 men and 114 women). During autopsy, left temporal (LT), right temporal (RT), and occipital (O) bone samples were extracted from each skull. Sample thickness (ST) was measured using multidetector computed tomographic imaging. The fracture load (FL) of each sample was measured by a bending test, in which the flexural strength (FS) was calculated. The FL and ST values for O were significantly greater compared with those of the LT and RT bones. The temporal bones were thinner compared with other parts of the skull and at greater risk for fracture. There is a need to take precautions to prevent temporal bone fractures. There were no significant differences in any of the values between LT and RT, indicating bilateral symmetry of the temporal bones. There were significant negative correlations between age and the FL and FS values for all sites in both sexes, except for O in the male samples, suggesting that older individuals are at increased risk for fractures. No significant correlations were observed between age and ST values in any of the samples. There were significantly positive correlations between FL and ST values at all sites regardless of sex.

Characteristics of indoor and outdoor fatal hypothermia cases in Chiba, Japan.

In forensic medicine, hypothermia is a frequently encountered cause of death, and this characteristic provides public health information to prevent similar deaths in the future. Previous studies revealed regional differences in hypothermia occurrence (indoors or outdoors). However, to our knowledge, no recent studies in Japan have directly compared the characteristics of indoor- and outdoor-onset cases based on forensic autopsy reports. Hence, this study aimed to determine the characteristics and risks of unexpected hypothermia-related death. It included 218 cases from the Chiba Prefecture, Japan, wherein forensic autopsies were performed and hypothermia was diagnosed; these cases were categorized into indoor- and outdoor-onset cases, and their characteristics were examined. The results showed no significant differences between the two groups in relation to the age of onset or residential environment (i.e., the presence or absence of cohabitants). The outdoor-onset group tended to have a higher incidence of dementia. Regarding the causes of hypothermia, the indoor group had more internal causes (p < 0.0001), whereas the outdoor group had more primary and external causes (p < 0.0001 and p = 0.0029, respectively). The indoor group was more undressed than the outdoor group. Atypical antipsychotic components were predominantly detected in the blood in the outdoor group (p = 0.0077). The body mass index tended to be lower in the indoor group than in the outdoor group. Broadening public awareness of the present study findings may aid in developing preventative strategies for hypothermia based on the location of onset.

Changes in SARS-CoV-2 viral load and titers over time in SARS-CoV-2-infected human corpses.

High viral titers of infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been detected in human corpses long after death. However, little is known about the kinetics of infectious SARS-CoV-2 in corpses. In this case series study, we investigated the postmortem kinetics of infectious SARS-CoV-2 in human corpses by collecting nasopharyngeal swab samples at multiple time points from six SARS-CoV-2-infected patients after their death. SARS-CoV-2 RNA was detected by quantitative reverse transcription-polymerase chain reaction from nasopharyngeal swab samples collected from all six deceased patients. A viral culture showed the presence of infectious virus in one deceased patient up to 12 days after death. Notably, this patient had a shorter time from symptom onset to death than the other patients, and autopsy samples showed pathological findings consistent with viral replication in the upper respiratory tract. Therefore, this patient died during the viral shedding phase, and the amount of infectious virus in the corpse did not decrease over time up to the date of autopsy (12 days after death). The findings of this study indicate that the persistence of SARS-CoV-2 in corpses can vary among individuals and may be associated with the stage of the disease at the time of death. These important results complement many previously reported findings on the infectivity of SARS-CoV-2 at postmortem.

Effect of ethanol on the diatom test using nitric acid or sodium hypochlorite.

OBJECTIVE: To study the quantitative effect of ethanol on the diatom test for water and lung samples. MATERIALS AND METHODS: In experiment 1, we tested 20 water samples taken from natural water areas. In experiments 1-1 and 1-2, each sample was digested with sodium hypochlorite (NaClO) solution (Purelox) and fuming nitric acid (HNO3), respectively. In Experiments 1-3 and 1-4, each sample was added to a lung sample containing few diatoms and digested with NaClO and HNO3, respectively. In Experiment 2, eight lung samples containing diatoms were digested with NaClO. Then, each digested sample was divided into two portions; one portion was washed with ethanol before being washed with water, and the other was washed only with water. After base-2 logarithmic transformation, the counts from the methods with and without ethanol wash were compared with a paired t-test. RESULTS: In experiments 1-1, 1-2, 1-3, 1-4, and 2, the geometric means of the ratios derived from the two methods (with/without ethanol) were 0.70 (95 % confidence interval [CI]: 0.65-0.77, P < 0.001), 0.83 (95 % CI: 0.73-0.93, P = 0.005), 3.00 (95 % CI: 2.31-3.91, P < 0.001), 0.91 (95 % CI: 0.79-1.04, P = 0.164), and 3.06 (95 % CI: 2.28-4.41, P < 0.001), respectively. CONCLUSION: Our experiments suggest that ethanol would be useless in diatom tests of water samples or in the conventional (HNO3) digestion of lung samples. However, ethanol is essential for the NaClO digestion of lung samples and could also be useful for other alternative methods of lung samples.

Relevance of diatom testing on closed organs of a drowned cadaver who died after receiving treatment for 10 days: A case report.

A male in his late adolescence fell into the sea and was found 50 min later with cardiopulmonary arrest. He was revived approximately 260 min after he drowned. Although he received several treatments, including venoarterial extracorporeal membrane oxygenation and continuous hemodiafiltration, he was clinically diagnosed with brain death. He died 10 days after the accident. The autopsy did not reveal any unremarkable findings other than those associated with post-resuscitation changes and medical treatment. The diatom test revealed 47.9, 311.6, and 577.5 diatom particles per gram from water, left lung, and right lung samples, respectively. No diatoms were detected in a 10 g liver sample, and 1 diatom was detected in each of approximately 12 g of bilateral kidney samples, which was different from the abundant species in the lung samples. The diatom test of the closed organs could be considered false negative for confirming drowning death since diatoms can also be detected in non-drowned cadavers on dry land. This suggests that diatoms might not reach the closed organs via circulation and that the diatom test of closed organ samples might no longer be necessary to confirm drowning deaths.

Age estimation by palatal suture using modified Kamijo's method.

The present study aimed to investigate the correlation between palatal suture obliteration and age in modern Japanese and to develop an age estimation equation by modifying Kamijo's (1949) method. The subjects were 195 Japanese skeletal remains (155 males and 40 females) whose age and sex were known. First, obliteration score (OS) was obtained by measuring palatal suture obliteration from photographic images taken at the time of forensic autopsy, and the correlation with age was examined; no significant correlation was found in females. Second, the palatal sutures were divided into 14 sections, and each section was scored from 0 to 4 points according to the degree of the suture obliteration. Suture scores (SS) were then calculated for each of the four sutures, and the sum of the 14 scores (TSS: total suture score) was used to perform regression analysis for age. For male and all subjects (male and female), age significantly increased (p < 0.001) according to increment of SSs for all sutures. TSS has the highest regression coefficient (r = 0.540), and the lowest standard error of estimation (13.54 years) for all of the patients. The intra- and inter-observer agreement scoring showed high reliability. Validation study using the formulae showed a high percentage of correct responses (80 %). In conclusion, age estimation regression formula by palatal suture using modified Kamijo's method was established for Japanese population, and the study showed the formula might be valid for age estimation.

Occupational accidental injury deaths in Tokyo and Chiba prefectures, Japan: A 10-year study (2011-2020) of forensic institute evaluations.

Occupational accidental injury deaths (OAIDs) are a major social problem, and the analysis of individual cases is important for developing injury prevention measures. In this study, OAIDs with autopsies performed at forensic facilities in the metropolitan area of Japan (Tokyo and Chiba prefectures) from 2011 to 2020 were reviewed. The epidemiological characteristics of these OAIDs (n = 136), which accounted for 13.5% of OAIDs reported in the region during the study period, were compared with those of non-occupational accidental injury deaths (non-OAID) cases (n = 3926). Among OAID cases, 134 (98.5%) were men and 13 (9.6%) were foreign-born workers, which was significantly more than in non-OAID cases (p < 0.001, respectively). OAIDs were most frequent in construction (39.0%) followed by the manufacturing category (21.3%). The percentage of OAIDs in workers aged 65 and over showed an increasing trend. Most accidents occurred just after the start of work or just before the workday ended, as well as during the peak months of the year. The most common type of accident was fall/crash from a height (25.0%), and the most common injury site was the chest; none of these cases were confirmed to have been wearing a safety belt properly. Among foreign-born workers, the most common type of accident was caught in/between. As the working population is expected to change in the future, and an increase in the number of older adults and foreign workers is expected, it is necessary to take preventive measures such as improving the work environment based on ergonomics and providing safety education.

Methamphetamine-related forensic autopsy cases in a Japanese prefecture over a 7-year period: Characteristics of deaths and blood concentrations.

Understanding the actual conditions of methamphetamine (MA)-related death is important from the perspectives of criminal justice and public health. In this report, we review 104 cases of MA-related death handled by our departments between January 2014 and December 2020. Based on information from police and autopsy examinations, we classified the cases into the following categories: "accidental intoxication" ("MA only" and "multiple drugs or alcohol"), "fatal disease" ("definitively MA-related," "possibly MA-related," and "unlikely MA-related"), "accident," "suicide," "homicide," and "undetermined." The total number and annual trends for each category and their respective femoral blood concentrations were investigated. "Fatal disease" was the most common category (48 cases), followed by "suicide" (25 cases), "accidental intoxication" (14 cases), and "accident" (11 cases). "Definitively MA-related" in which MA may have played a role in their onset or exacerbation accounted for the majority of "fatal disease": 12 cases of heart disease, 4 cases of aortic dissection, 12 cases of cerebral hemorrhage, and 4 cases of subarachnoid hemorrhage. Cases classified as "definitively MA-related" died with lower femoral blood concentrations of MA compared with "MA only." Cases with "fatal disease" might have been misdiagnosed as "death by natural causes" if a proper autopsy and toxicology examinations were not performed. In death investigations, it is necessary to keep in mind that there are some MA-related deaths, and efforts should be made to increase awareness about the risk of death in using this drug.

COVID-19 Analysis in Tissue Samples Acquired by Minimally Invasive Autopsy in Out-of-Hospital Deaths with Postmortem Degeneration.

Minimally invasive autopsy (MIA) is an alternative to a full autopsy for the collection of tissue samples from patients' bodies using instruments such as a biopsy needle. MIA has been conducted in many cases of coronavirus disease 2019 (COVID-19) and has contributed to the elucidation of the disease pathogenesis. However, most cases analyzed are hospital deaths, and there are few reports on the application of MIA in out-of-hospital deaths with varying extents of post-mortem changes. In this study, MIA and autopsies were performed in 15 patients with COVID-19 2-30 days after death, including 11 out-of-hospital deaths. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome detection by reverse transcriptase quantitative polymerase chain reaction using MIA samples was mostly consistent with autopsy samples, particularly lung tissue, even in out-of-hospital cases. MIA had high sensitivity and specificity (> 0.80). Histological examination of lung tissue obtained by MIA showed characteristics of COVID-19 pneumonia, with 91% agreement with autopsy samples, whereas localization of SARS-CoV-2 protein in lung tissue was indicated by immunohistochemistry, with 75% agreement. In conclusion, these results suggest that MIA is applicable to out-of-hospital deaths due to COVID-19 with various postmortem changes, especially when autopsies are not available.

High titers of infectious SARS-CoV-2 in corpses of patients with COVID-19.

OBJECTIVES: The prolonged presence of infectious SARS-CoV-2 in deceased patients with COVID-19 has been reported. However, infectious virus titers have not been determined. Such information is important for public health, death investigation, and handling corpses. The aim of this study was to assess the level of SARS-CoV-2 infectivity in the corpses of patients with COVID-19. METHODS: We collected 11 nasopharyngeal swabs and 19 lung tissue specimens from 11 autopsy cases with COVID-19 in 2021. We then investigated the viral genomic copy number by real-time reverse transcription-polymerase chain reaction and infectious titers by cell culture and virus isolation. RESULTS: Infectious virus was present in six of 11 (55%) cases, four of 11 (36%) nasopharyngeal swabs, and nine of 19 (47%) lung specimens. The virus titers ranged from 6.00E + 01 plaque-forming units/ml to 2.09E + 06 plaque-forming units/g. In all cases in which an infectious virus was found, the time from death to discovery was within 1 day and the longest postmortem interval was 13 days. CONCLUSION: The corpses of patients with COVID-19 may have high titers of infectious virus after a long postmortem interval (up to 13 days). Therefore, appropriate infection control measures must be taken when handling corpses.