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Forensic entomology (FE) involves the collection and analysis of necrophagous insects and arthropods for postmortem interval estimation based on their growth and succession. In Japan, research on FE is limited, and the predominant species of necrophagous insects and their distribution have not been clarified. The present study aimed to clarify the actual situation of insects, targeting Diptera collected from human cadavers, in Chiba Prefecture. We targeted the cases for which specimens could be collected from forensic autopsies conducted at the Legal Medicine Department of Chiba University Graduate School of Medicine or from human bodies handled by the Chiba Police in 2019. We specifically chose cases from which adult insects could be bred. Fifty cases were applicable, and 47 cases were successful in rearing adult flies. Sixteen species of Diptera were identified: six species of Calliphoridae, four species of Sarcophagidae, and a few species of Fanniidae, Muscidae, Phoridae, Piophilidae, and Stratiomyidae. The most frequently observed species were Lucilia sericata (20 cases) and Chrysomya pinguis (18 cases). For the first time, globally, we recorded the presence of Fannia prisca in a human cadaver. In conclusion, this is the first report to reveal the presence of carrion flies in human death cases in Chiba Prefecture, Japan. Research in FE is important in the context of the Japanese natural and social environment since this can aid forensic investigations conducted by authorities.

Estimation of age at death is important in forensic investigations of unknown remains. There have been several reports on applying the degree of osteophyte formation—an age-related change in the vertebral body—for age estimation; however, this method is not yet established. This study investigated a method for age estimation of modern Japanese individuals using osteophytes measured on CT images. The sample included 250 cadavers (125 males) aged 20–95 years. The degree of osteophyte formation was evaluated as score O (0–5 points), and the degree of fusion of the osteophytes between the upper and lower vertebrae was evaluated as score B (0–2 points). Age estimation equations were developed using regression analyses with seven variables, determined by scores O and B, and the equation with the smallest standard error of estimate (SEE) was obtained when the number of vertebrae with score O ≥ 2 was used as the explanatory variable. Age estimation with SEE of about 10 years was possible even when partial vertebrae with a high degree of osteophyte formation were used, showing its potential for practical application. The cutoff value for age estimation was established using the receiver operating characteristic curve analysis, wherein good results were obtained for all variables (area under the curve ≥ 0.8). The combination of the estimation equation and the cutoff value can narrow the range of age estimates.

BackgroundThe prolonged presence of infectious severe acute respiratory syndrome coronavirus (SARS-CoV-2) in deceased coronavirus disease 2019 (COVID-19) patients has been reported. However, infectious virus titers have not been determined. Such information is important for public health, death investigation, and handling corpses.AimThe aim of this study was to assess the level of SARS-CoV-2 infectivity in COVID-19 corpses.MethodsWe 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.ResultsInfectious virus was present in 6 of 11 (55%) cases, 4 of 11 (36%) nasopharyngeal swabs, and 9 of 19 (47%) lung specimens. The virus titers ranged from 6.00E + 01 plaque-forming units (PFU)/mL to 2.09E + 06 PFU/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.ConclusionCOVID-19 corpses 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.

To study the quantitative effect of ethanol on the diatom test for water and lung samples.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 (HNOIn 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.Our experiments suggest that ethanol would be useless in diatom tests of water samples or in the conventional (HNO

Acupuncture is practiced as a complementary medicine worldwide. Although it is considered a safe practice, pneumothorax is one of its most common serious complications. However, there have been few reports of deaths due to pneumothorax after acupuncture treatment, especially focused on electroacupuncture. We report an autopsy case of a man in his 60s who went into cardiopulmonary arrest and died immediately after receiving electroacupuncture. Postmortem computed tomography (PMCT) showed bilateral pneumothoraces, as well as the presence of numerous gold threads embedded subcutaneously. An autopsy revealed two ecchymoses in the right thoracic cavity and a pinhole injury on the lower lobe of the right lung, suggesting that the needles had penetrated the lung. There were marked emphysematous changes in the lung, suggesting that rupture of bullae might also have contributed to bilateral pneumothoraces and fatal outcome. The acupuncture needles may have been drawn deeper into the body than at the time of insertion due to electrical pulses and muscle contraction, indicating the need for careful determination of treatment indications and technical safety measures, such as fail-safe mechanisms. This is the first case report of fatal bilateral pneumothoraces after electroacupuncture reported in the English literature. This case sheds light on the safety of electroacupuncture and the need for special care when administering it to patients with pulmonary disease who may be at a higher risk of pneumothorax. This is also the first report of three-dimensional reconstructed PMCT images showing the whole-body distribution of embedded gold acupuncture threads, which is unusual.

Ropinirole is an antiparkinsonian drug and has recently been suggested to be effective in amyotrophic lateral sclerosis. It is expected that ropinirole prescriptions will increase in the near future. However, the fatal concentration in blood is unclear at this time. Therefore, we report a fatal case involving ropinirole intoxication and discuss the fatal concentrations with reference to several autopsy cases involving ropinirole. Ropinirole was quantified in femoral vein blood, cardiac blood, and urine from five autopsy cases in which ropinirole was detected by drug screening in our laboratory. One is a ropinirole intoxication case (this report) and the others were non-intoxication cases. Their ropinirole concentrations were compared and discussed. The ropinirole concentration in this case was 100 ng/mL in femoral blood, 160 ng/mL in cardiac blood, and 1840 ng/mL in urine. The ropinirole concentrations in the four non-ropinirole poisoning cases were 7–35 ng/mL (mean: 24 ng/mL) in femoral blood, 13–100 ng/mL (mean: 60 ng/mL) in cardiac blood, and 140–1090 ng/mL (mean: 640 ng/mL) in urine. Cardiac/peripheral ratios were in the range of 1.6–2.1 (mean 1.8). There were no obvious signs of overdose, and the high cardiac/peripheral blood ratio suggested that postmortem redistribution may have occurred, but the peripheral blood ropinirole concentration (100 ng/mL) was obviously higher than that reported in the previous fatal case of ropinirole poisoning (64 ng/mL). Based on these results, the cause of death in this case was considered to be shock and fatal arrhythmia due to ropinirole poisoning. This case provides important data on postmortem blood and urinary levels of ropinirole poisoning.

Ludwig's angina is characterized by inflammation of the sublingual and submandibular spaces and is mainly caused by odontogenic infection, which leads to cellulitis of the soft tissues of the floor of the mouth and the neck. This causes asphyxia due to elevation and posterior deviation of the tissues of the floor of the mouth. We report a fatal case of airway obstruction due to Ludwig's angina. A woman in her forties who had no physical complications, but had a mental illness, was undergoing outpatient dental treatment for caries in the first premolar of the left mandible. She was admitted to a psychiatric hospital because of insomnia caused by pain, where she developed cardiopulmonary arrest while sleeping and died 14 days after onset of the dental infection. Postmortem computed tomography (PMCT) prior to autopsy showed swelling of the soft tissues-from the floor of the mouth to the oropharyngeal cavity, the supraglottic larynx, and the prevertebral tissue. Autopsy revealed a markedly swollen face and neck, an elevated tongue, and a highly edematous epiglottis and laryngopharyngeal mucosa. There was also cellulitis and abscess of the facial, suprahyoid, and neck musculature, which suggested that the cause of death was asphyxiation due to airway obstruction. This was an alarming case, with mental illness leading to risk of severe odontogenic infection, and in which obesity and use of antipsychotic medication might have acted synergistically leading to airway obstruction. This is also a case of Ludwig's angina captured by PMCT, which has rarely been reported.

In Japan, a new cause-of-death investigation system and related new laws were enacted in the mid-2010s. These laws provided for an autopsy system for non-criminal unnatural deaths and a medical accident investigation system outside the criminal justice process for health care-related deaths. We retrospectively explored changes in the number and characteristics of medico-legal autopsy cases of health care-related deaths in Chiba Prefecture, Japan, and examined trends over time during these reforms. We found that the percentage of forensic autopsies based on the Code of Criminal Procedure for health care-related deaths had decreased significantly. The number of autopsies of accidental and unintentional deaths in nursing homes, which are not covered by the newly established medical accident investigation system, has been increasing, reflecting the ageing of society. The trend toward decriminalisation of health care-related deaths was expected to contribute more to medical safety if the scope was expanded and a system for disclosure of autopsy information was established.

To statistically clarify the prevalence and risk factors of infections in forensic autopsy cases in Chiba Prefecture, Japan. The aim was to improve preventive measures against infection in forensic autopsies.We retrospectively investigated the positive detection rates of five infections (hepatitis B, HBV; hepatitis C, HCV; human immunodeficiency virus, HIV; human T-lymphotropic virus, HTLV; Treponema pallidum, TP) using 1491 samples obtained in forensic autopsy at our facility from 2014 to 2018. In addition, risk factors related to infection such as methamphetamine and tattoos were analyzed. Pearson's chi-square test was used for statistical analysis, and the difference was judged to be significant at p 0.05.Among our samples, 9.0% of cadavers tested positive for infection, and the prevalence rates for HBV, HCV, HIV, HTLV, and TP were 1.0%, 6.7%, 0.3%, 0.7%, and 1.1% respectively. Statistically, cadavers linked to information about methamphetamine use had a 7.2 times higher rate of infection, and those with tattoos had a 5.6 times higher rate of infection, with HCV being the predominant cause.To limit the risk of infection among autopsy workers, cadavers and samples should be handled on the presupposition that the bodies are at risk of infections. It is also important to obtain as much information as possible about the medical history and potential illegal drug use to help assess the risk of infection in a patient during forensic autopsy. We propose that all autopsy cases should be screened for infections whenever possible.

Purpose Lisdexamfetamine (LDX), which is used for the treatment of attention-deficit/hyperactivity disorder and narcolepsy, is composed of l-lysine attached to dextroamphetamine (d-amphetamine). In this article, we report a forensic autopsy case in which prescription drugs were unknown at autopsy. While amphetamine was detected, methamphetamine could not be detected by liquid chromatography–tandem mass spectrometry (LC–MS/MS) in any of samples collected. Thus, we aimed to quantify LDX concentrations in autopsy samples and to prove that the amphetamine detected in this case was due to metabolized LDX. Methods Femoral vein blood, cardiac whole blood, urine, and gastric content samples were taken at autopsy for toxicological analysis. Qualitative and quantitative analyses were performed using LC–MS/MS. In addition, optical isomer separation for the amphetamine detected was conducted. The stability of LDX in whole blood and urine was also examined at three different temperatures. Results The concentrations of LDX were d-amphetamine by optical isomer separation. The d-amphetamine detected was considered to be derived from LDX. Stability experiments revealed that LDX in whole blood decreased at ambient temperature. Conclusions The results in the present case report may be useful in interpreting whether or not the amphetamine detected in a cadaver is a metabolite of LDX.

The diatom test method using sodium hypochlorite (NaClO) was equivalent to the conventional method in water samples. However, the method using NaClO was inferior to the conventional method in lung samples, in which ethanol was used and the reaction with NaClO was longer compared with the method in water samples. Using water samples, we aimed to clarify whether these differences affect the diatom test result.Thirteen water samples from natural water sources were each divided into four parts corresponding to four (2 × 2) digestion methods: 3 "digestion" vs. 1 "digestion" and with ethanol vs. without ethanol. After the base-2 logarithmic transformation, the diatom counts were analyzed using three-way analysis of variance (ANOVA); factor 1 was "digestion times," factor 2 was "ethanol," and factor 3 was "sample number," and the interaction between factors 1 and 2 was also analyzed.The geometric means of the diatoms from the 3 "digestion" with ethanol method, the 3 "digestion" without ethanol method, the 1 "digestion" with ethanol method, and the 1 "digestion" without ethanol method were 373.5, 551.8, 436.6, and 522.0, respectively. ANOVA showed a significant difference in factor 2 (P = 1.7 × 10Ethanol may decrease the diatom count in the diatom test using NaClO. In contrast, the diatom frustules do not dissolve through three-times digestion using NaClO.

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.

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.

Postmortem computed tomography (PMCT) images help identify individuals and extract information from corpses. PMCT may substitute for a standard examination when bodies are severely damaged or when resources are limited in a mass fatality incident. In such situations, the dental information revealed by PMCT has the potential to narrow down candidates for identification further. However, the validity of the dental findings obtained from PMCT images remains unclear. In this study, we investigated the validity of dental findings on PMCT images compared to regular dental examinations as the reference standard. We routinely collected PMCT images of 148 unidentified corpses and compared the dental findings of each tooth obtained from PMCT with those of a regular dental examination. The validity of dental findings of PMCT was measured by sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). We also evaluated the accuracy of dental findings per corpse using 32 teeth as the denominator. The dental findings with high sensitivity and high specificity were a present tooth (0.96 and 0.97), a missing tooth (0.97 and 0.97), pontic (0.92 and 1.00), a dental implant (1.00 and 1.00), and a root filling (0.94 and 0.99). The mean accuracy of the dental findings per corpse in PMCT was 95.6% (standard deviation: 6.9, minimum ≤ median ≤ maximum: 65.6 ≤ 100 ≤ 100). The number of corpses with 100% accuracy in the dental findings was 81 (54.7%). The information obtained in this study highlights the potential use of PMCT during human identification in several settings with limited resources, such as the number of specialists present and the condition of the corpses.

Cyanide is a powerful and rapidly acting poison. In Japan, cyanide poisoning is rare, and regular cyanide testing can be costly and time consuming. In contrast, alcohol analysis is routinely performed in most forensic laboratories. In this study, we attempted to develop a method for the simultaneous quantification of cyanide and alcohols in blood using headspace gas chromatography (HS-GC). As nitrogen-phosphorus detection (NPD) is more sensitive to hydrogen cyanide than mass spectrometry (MS), a Deans switch was used to switch the detectors during a single run. The separation provided by three analytical columns, PoraBOND Q, CP-Sil 5 CB, and HP-INNOWax, was investigated, and PoraBOND Q was selected. The use of HS-GC-MS/NPD with a Deans switch enabled the simple and simultaneous quantification of cyanide, ethanol, and 1-propanol. Eighteen other volatile compounds were detected in the SIM/scan mode of the MS.

We report a fatal case of airway obstruction caused by spontaneous retropharyngeal hematoma (RH) in the setting of treatment with dipyridamole. A 90-year-old woman presented with cervical swelling, neck and chest ecchymoses, and complaints of dyspnea. She suffered cardiopulmonary arrest in the ambulance, and her death was confirmed after transportation to the hospital. The major finding of postmortem computed tomography (PMCT) prior to autopsy was widening of the prevertebral soft tissue. The results of the autopsy indicated that the cause of death was mechanical asphyxia, secondary to pharyngeal and laryngeal compression caused by the RH. There were no evident injuries, medical interventions, or particular diseases, suggesting the spontaneous cause of the RH. To the best of our knowledge, this is the first report of a fatal case secondary to spontaneous RH that was revealed through PMCT imaging.

To elucidate postmortem computed tomography (PMCT) and postmortem magnetic resonance (PMMR) imaging findings suggesting massive fat embolism. 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. 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. 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.

To estimate forensic age, the Study Group of Forensic Age Diagnostics of the German Association of Forensic Medicine (AGFAD) has recommended the radiological investigation of the medial clavicular epiphysis (MCE). This study aimed to investigate the relation between the maturation of the MCE and age in a contemporary Japanese population using a standard five-stage system by Schmeling et al. and Kellinghaus substage system. A total of 207 cadavers (128 males, 79 females) of known age and sex that underwent postmortem computed tomography (CT) and subsequent forensic autopsy were obtained. The earliest appearance for stage 3c was 18.1 years in the male subjects and 18.4 years in the female subjects. Our results were consistent with those in previous studies and indicate that stage 3c is crucial for medico-legal processes in evaluating age in various countries. Similar studies on the staging of the MCE using CT images in other populations, particularly in Asians, are required.

Estimation of age at death is important in forensic investigations of unknown remains. There have been several reports on applying the degree of osteophyte formation-an age-related change in the vertebral body-for age estimation; however, this method is not yet established. This study investigated a method for age estimation of modern Japanese individuals using osteophytes measured on CT images. The sample included 250 cadavers (125 males) aged 20-95 years. The degree of osteophyte formation was evaluated as score O (0-5 points), and the degree of fusion of the osteophytes between the upper and lower vertebrae was evaluated as score B (0-2 points). Age estimation equations were developed using regression analyses with seven variables, determined by scores O and B, and the equation with the smallest standard error of estimate (SEE) was obtained when the number of vertebrae with score O ≥ 2 was used as the explanatory variable. Age estimation with SEE of about 10 years was possible even when partial vertebrae with a high degree of osteophyte formation were used, showing its potential for practical application. The cutoff value for age estimation was established using the receiver operating characteristic curve analysis, wherein good results were obtained for all variables (area under the curve ≥ 0.8). The combination of the estimation equation and the cutoff value can narrow the range of age estimates.

Background In forensic medicine, the diatom test is used to diagnose drowning. Drowning and postmortem immersion can be distinguished by calculating the ratio of diatom concentration in the lungs and drowning water (L/W ratio). However, this claim was based on the unproven hypothesis that diatoms may be concentrated in the lungs due to respiratory movements. This study was conducted to examine whether the L/W ratio increased with experimental water injection. Methods A total of two experiments was performed using 22 non-drowned cat carcasses found on dry land. First, for the experimental postmortem immersion, we soaked seven whole-body cat carcasses in pond water for an hour. Second, the pond water was experimentally injected one or ten times into each harvested lung from seven and eight cats, respectively. In the diatom test, two diatom species (Aulacoseira ambigua and Discostella asterocostata) that were dominantly observed in pond water as well as other diatom species were counted separately. The L/W ratios of each cat were calculated. Univariate linear regression analysis was performed to demonstrate the association among L/W ratios and the three categories of the experiments. The L/W ratios of the two experiments were compared with those of drowning or postmortem immersion cases of humans or cats. Results It was revealed that the clear L/W ratio differences between the three groups (experimental postmortem immersion 0.9 for all diatom counting) were with statistically significant as proven by the univariate regression analyses. In actual cases of cats and humans, L/W ratios were>0.4 for drowning and Conclusion The L/W ratio increased with multiple experimental water injections into the lungs, thereby verifying the validity of the diatom concentration test to diagnose drowning. The diatom test can be used to distinguish between drowning and postmortem immersion in humans and cats by calculating the L/W ratio.

The diagnosis of drowning is an important issue in forensic investigations. Moreover, discriminating between seawater and freshwater drowning is crucial to identify where the drowning occurred. The present study aimed to investigate electrolyte concentrations in pleural fluid in decomposed bodies in late postmortem intervals and derive cut-off values for the diagnosis of seawater and freshwater drowning.Data were collected from 44 seawater drowning cases, 60 freshwater drowning cases, and 30 non-drowning cases with pleural effusion which served as controls. The levels of sodium ion (NaThe receiver operating characteristic analysis revealed that the sensitivity and specificity were both 1.000 for SUMThe electrolyte concentrations in pleural effusion may be useful for the diagnosis of drowning in decomposed bodies with a longer postmortem interval.

Streptococcal toxic shock syndrome (STSS) is a severe infection most commonly caused by group A streptococcus. It is clinically characterized by rapidly progressive multiple organ failure and septic shock. This report presents four fatal cases associated with STSS. In two cases (cases 2 and 3), the portals of entry may be ulcer and mucosa at the surgical site; the initial symptoms in these cases included fever. In the other cases, the portal of entry was unknown; the initial symptom was pain. In two cases (cases 1 and 3), malpractice was suspected before autopsy. At autopsy, blood culture was positive for group A streptococcus in all of the present cases. Although C-reactive protein levels were increased, procalcitonin levels were not markedly elevated. This is the first report of autopsy cases associated with STSS in which postmortem computed tomography was performed; the swelling of muscles and increased concentrations of peripheral subcutaneous tissue without gas may be characteristic findings. Histology revealed extensive bacterial colonies and necrosis with mild neutrophilic reaction in two cases (cases 3 and 4), and hemophagocytosis in two cases (cases 1 and 4). It is essential to perform appropriate examinations and make a proper diagnosis because STSS still has a high fatality rate and medical malpractice is often suspected. In addition, it is important to evaluate STSS again for the reduction of the fatal cases associated with STSS.

In recent years, there has been an increase in the use of phosphodiesterase type 5 inhibitors (PDE5i) that are purchased from abroad without a doctor’s diagnosis via the Internet or other means. We report six cases in which nonprescription use of PDE5i may have led to death. Among the four deceased individuals who were believed to have experienced sudden cardiac death, three (cases 1–3) had a history of cardiovascular disease, which is a contraindication, and the remaining case (case 4) involved combined use of multiple PDE5i. Sildenafil (0.063 µg/mL, 0.087 µg/mL) was detected in two of the four cases of sudden cardiac death. Tadalafil (0.096 µg/mL) was detected in one of the remaining two cases, and tadalafil (0.197 µg/mL) and vardenafil (0.011 µg/mL) were detected in the other case. Sildenafil (0.032 µg/mL), tadalafil (0.062 µg/mL), and ethanol were detected in a traffic accident case with a history of contraindications. In a case of asphyxiation by vomit aspiration, autopsy showed 90% stenosis in the anterior descending branch of the coronary artery, and sildenafil (0.063 µg/mL) was detected. To the best of our knowledge, this is the first report of postmortem blood levels of tadalafil and vardenafil likely contributing to the cause of death. Despite all the warnings about the dangers of using PDE5 inhibitors, cases of PDE5i contributing to death are still identified during autopsies. Therefore, raising public awareness of the risks of the risks associated with the imported drug use by individuals is necessary.

The purpose of this study was to assess the incidence of fatal hemorrhage complicated with methamphetamine (MA) poisoning and to examine the postmortem computed tomography (PMCT) features of fatal intracerebral hemorrhage (ICH) with and without MA poisoning. The study also attempted to determine the differences in PMCT between those two groups. Consecutive medicolegal autopsy data from November 2011 to February 2018 were searched for 3044 cases. First, the incidence and distribution of all cases of nontraumatic fatal hemorrhage with various causes were examined. Second, cases of ICH on the basal ganglia and brain stem were extracted. The PMCT findings were compared with respect to nine parameters: volume of hematoma, ventricular perforation, midline shift distance, aortic calcification, calcification of aortic valve, calcification of coronary artery, cardiothoracic ratio, circumference of ascending aorta, and volume of bladder contents. Of the 3044 cases, 97 were nontraumatic fatal hemorrhage; of these 97 cases, 20 were classified as MA poisoning with 9 ICH cases, and 60 cases were classified as non-MA poisoning with 14 ICH cases. A statistically significant difference in ages was observed between the two groups. On PMCT comparison of ICH, statistically significant differences were evident in the midline shift distance and calcification of the aortic valve. Forensic radiologists should be aware of the possibility of ICH with MA poisoning if fatal hemorrhage is detected on PMCT. Younger age, less calcification of the aortic valve, and a remarkable midline shift may be the keys to recognition.

A 5-year-old boy with a chromosome-9 abnormality and multiple external and visceral malformations was found in cardiopulmonary arrest during a regular visit to the hospital; he did not respond to cardiopulmonary resuscitation and died. An odontoid process fracture and calcification and fibrosis of the muscles around the superior cervical vertebra were observed during the autopsy. Postmortem computed tomography revealed an anterior dislocation of the atlas; odontoid synchondrosis fracture; and delayed, incomplete bony fusion of the odontoid process relative to his age. The cause of his death was a superior spinal cord injury. The tissue surrounding the upper cervical spine presented with myositis ossificans, suggesting a prior injury. He experienced a minor traffic accident 3 months before his death. It was concluded that the odontoid synchondrosis fracture occurred during the accident based on the incomplete bony fusion and atlantoaxial instability, which were consistent with the findings of myositis ossificans. Delayed fatal dislocation may then have occurred under the influence of a minor external force. Odontoid process abnormalities and atlantoaxial instability are common in patients with trisomy 21 and other congenital diseases; however, the condition's association with chromosome-9 abnormalities has not been reported. In children with various chromosomal abnormalities, periodic assessment of instability and morphology of the cervical spine, and a lowered examination threshold for the children at risk, could prove useful in the prevention injuries leading to fatality, and provide additional information to rule out abuse.

Atmospheric radiocarbon (14C) levels increased from 1955 to 1963 due to atmospheric nuclear weapon tests, and then decreased. As 14C accumulates in human tooth enamel while the enamel is being formed, 14C can be used to estimate the birth year of unidentified bodies. Measurement results of 14C content in tooth enamel using accelerator mass spectrometry vary depending on the enamel's sample site. To address this problem, a method for equalizing samples using a pulverizer was considered in this study. Regarding the tube and cone used as the pulverizer, (1) a polycarbonate tube and stainless steel cone, (2) a stainless steel tube and cone, and (3) a tungsten carbide tube and cone, were compared. In (1), the modern carbon ratio was approximately half that of the normal ratio of 100 pMC, with which accurate dating was impossible, and in (2), a high background value was obtained for IAEA-C1, which was pulverized using a reusable tube and cone. In (3), the 14C content for IAEA-C1 pulverized using reusable tube and cone, which was washed with quartz sand, was 0.31 ± 0.01 pMC. This result did not show any problems regarding background value. Therefore, the use of tungsten carbide products and washing with quartz sand is recommended for 14C measurement of pulverized teeth.

We report a case of fatal olanzapine-induced ketoacidosis in which pneumomediastinum (PM) and subcutaneous emphysema (SE) were detected on postmortem computed tomographic (CT) images. A man in his forties was found in a state of cardiopulmonary arrest with profuse perspiration, and 50 empty capsules of olanzapine (10 mg) and flunitrazepam (1 mg) were found in his room. The major findings of postmortem CT prior to autopsy were PM and SE from the lower half of the face to the height of the first rib. The results of autopsy, biochemical tests, and toxicological analyses indicated the cause of death to be fatal ketoacidosis induced by olanzapine intoxication. No injuries, medical interventions, or particular diseases were evident, suggesting that PM and SE were caused by ketoacidosis. Our findings indicated that toxicological analyses should be performed when PM and SE are detected on CT images

Animal studies suggested that halogenated hydrocarbons such as 1,1-difluoroethane (DFE) sensitized myocardial tissues to catecholamines and might cause fatal arrhythmia. In this paper, we report a case of a fatality that was associated with DFE abuse, and quantified DFE concentrations in postmortem specimens using gas chromatography–mass spectrometry (GC–MS). Femoral vein blood, cardiac blood, and urine samples were taken from the autopsy for toxicological analysis. We have established a detailed procedure for quantification of DFE in human blood and urine by GC–MS and have presented its validation data. The concentrations of DFE in this case were 481, 591 and 201 µg/mL in femoral vein blood, cardiac blood and urine samples, respectively, which were much higher than those in previous cases measured by gas chromatography–flame ionization detection. Thus, in the absence of other remarkable autopsy findings, the cause of death was determined to be DFE intoxication. To the best of our knowledge, this is the first case report of quantification of DFE in human blood and urine specimens by GC–MS.

Aim: We examined the possibility of tracing the origins of cadavers based on the geographic distribution of genotypes of the latent human herpes simplex virus type 1 (HSV-1). Materials & methods: PCR of the V1 region (667 bp) was performed using DNA from human trigeminal ganglia of 107 Japanese cadavers. The phylogenetic analysis was performed using 16 cadavers in Chiba district and 84 reference sequences in the world. Results: The HSV-1 DNA detection rate was 54.2% and it increased with age. This phylogenetic tree showed four large clusters, types I–IV (African, European, Asian and the mixed-type), and two subclusters (Japanese–Korean type and Japanese–Chinese type) in type III. Conclusion: HSV-1 genotype is possible to trace the geographical origin of unidentified cadavers.

Analysis of ions in sphenoidal fluid can be a useful index for the diagnosis of drowning. We evaluated the reference ranges of non-drowning cases using statistical methods and three indices: sodium ion (Na+), summation of sodium and potassium ions (SUMNa+K), and summation of sodium, potassium and chloride ions (SUMNa+K+Cl). The reference ranges were 96≤Na+

Rapid and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in dead bodies is essential to prevent infection among those working with dead bodies. This study focused on the Smart Amplification (SmartAmp) method, which has a short examination time (approximately an hour), is simple to perform, and demonstrates high specificity and sensitivity. This method has already been used for clinical specimens; however, its effectiveness in dead bodies has not been reported. This study examined the SmartAmp method using 11 autopsies or postmortem needle biopsies performed from January to May, 2021 (of these, five cases tested positive for SARS-CoV-2 by quantitative real-time polymerase chain reaction (qRT-PCR) and six cases tested negative). Swab samples were collected from the nasopharynx, oropharynx, or anus and the SmartAmp and qRT-PCR results were compared. For the nasopharynx and oropharynx samples, the same results were obtained for both methods in all cases; however, for the anal swabs, there was one case that was positive according to qRT-PCR but negative according to the SmartAmp method. The SmartAmp method may therefore be less sensitive than qRT-PCR and results may differ in specimens with a low viral load, such as anal swabs. However, in the nasopharynx and oropharynx specimens, which are normally used for testing, the results were the same using each method, suggesting that the SmartAmp method is useful in dead bodies. In the future, the SmartAmp method may be applied not only during autopsies, but also in various situations where dead bodies are handled.

OBJECTIVE To determine the efficacy of using sodium hypochlorite (NaClO, Purelox) as a simple and rapid alternative digestion method for the diatom test through a quantitative comparison with the conventional method that uses fuming nitric acid (HNO3). MATERIALS AND METHODS In experiment 1, using 30 water samples, the NaClO and HNO3 methods were compared using paired t-test. In experiments 2 and 3, we included blank human lung plus 13 water samples and total of 32 drowned human lung samples, respectively, to compare the NaClO and HNO3 methods using paired t-test. The relationship between the concentration ratio and background factors was tested in experiment 3. Welch's t-test was used to determine differences in the ratio between the lung side and sex, whereas Pearson's correlation coefficient was used to determine the correlation between the ratio and either age or postmortem interval. The geometric mean of two counts was used for each specimen and all counts were logarithmically transformed to base 2 in the statistical analysis. RESULTS The NaClO method was completed within 80 min for any sample. In experiment 1, there was no significant difference between the NaClO and HNO3 methods using water samples (the mean of the ratios: 0.99, 95% confidence interval (95%CI: 0.89-1.10, P = 0.80). In experiment 2, the count of the NaClO method was lower than that of the HNO3 method using lung plus water samples (the mean of the ratios: 0.47, 95% CI: 0.35-0.65, P = 0.0002). In experiment 3, the concentration of the NaClO method was lower than that of the HNO3 method using drowned lung samples (the mean of the ratios: 0.28, 95% CI: 0.20-0.38, P

To clarify the differences between postmortem CT (PMCT) and autopsy findings in the postmortem detection of cervical spine injuries (CSIs).Our department's forensic pathology database was searched for CSI cases. In each case, the autopsy data and radiologists' interpretations were reviewed for the presence of bone fractures and intervertebral injuries.The study included 42 cases. For both bone fractures and intervertebral injuries, no substantial concordance between PMCT and autopsy findings was observed (McNemar's test: p0.001 and p0.001, respectively). Regarding bone fractures, more injuries were detected with CT than with autopsy (CT: 74, autopsy: 23). The percentage of CT-detected fractures that were missed at autopsy (77.0%, 57/74) was higher than the percentage of autopsy-detected fractures missed with CT (26.1%, 6/23). Regarding intervertebral injuries, fewer injuries were detected with CT than with autopsy (CT: 40, autopsy: 80). The percentage of CT-detected injuries that were missed at autopsy (35.0%, 14/40) was lower than the percentage of autopsy-detected injuries that were missed with CT (67.5%, 54/80).A substantial number of CSIs were detected by either PMCT or autopsy alone. Accurate evaluation for CSI requires the use of both methods.

Sex determination is an integral and foremost step in determining the identity of an unknown individual. The present study aimed to examine skeletal sexual dimorphism of the sacrum and coccyx using computed tomography (CT) images in a contemporary Japanese population and to derive discriminant function formulae for sex determination. Data were collected from 230 cadavers (115 male and 115 female subjects) subjected to postmortem CT scanning and subsequent forensic autopsy. Seven measurements of the sacrum and coccyx were obtained from CT images of each subject. The measurements were analyzed using descriptive statistics and discriminant function analyses. All except one measurement exhibited statistically significant sexual dimorphism, and the maximum sex determination rate was 83.5% according to discriminant function analyses. The results of this study suggest that discriminant analysis of sacral and coccygeal traits may be useful for the sex determination of skeletal remains in the Japanese population when applied with additional methods, such as morphological trait evaluation of other available bones.

Age estimation of cadavers from post-mortem “chest plate” using conventional radiography, which involves radiographic assessment of ossification around the sternum and rib ends, has been evaluated without fruitful results. This study examined the value of images of the chest plate obtained by three-dimensional post-mortem CT for estimation of age at time of death in a Japanese population. Five chest plate ossification scores were evaluated in 320 subjects, including ossification of the first costal cartilage (OF), ossification of the second to seventh costal cartilages at the rib (OR) and sternal (OS) ends, fusion of the manubriosternal joint (FM), and fusion of the xiphisternal joint (FX). OS was found to have the highest correlation with age while FM had no significant correlation. The best composite score for age estimation was the summative score for both sides of the OS and the right side of the OF and FX, for which the coefficient of determination (R2) and the standard error of estimation (SEE) were 0.608 and 12.44 years, respectively, for men and 0.590 and 14.65 years for women. The accuracy of the model was tested in a further 26 male and 24 female subjects, and the accuracy rate within the first SEE was 57.69% and 70.83%, respectively. This rapid and non-invasive method of age estimation in the chest plate area is superior to conventional methods and could be useful for estimation of age at time of death in the Japanese population.

Purpose The purpose of this study was to determine the incidence of three-rooted mandibular first molars in a contemporary Japanese population using multidetector computed tomography (MDCT) and examine whether this characteristic root form may be useful for identification purposes. Methods Prior to their forensic autopsies, we obtained MDCT scans of 365 cadavers (255 males, 110 females) with mandibular first molars on both sides. Altogether, 730 mandibular first molars were examined morphologically on reconstructed CT images, and the incidence of three-rooted molars was recorded. The results were analyzed statistically to determine sex and left–right differences using the chi-square test. Results In all, 189 (25.9%) of the 730 mandibular first molars had three roots: 79 (31.0%) in the 255 males and 35 (31.8%) in the 110 females. No statistically significant difference was found between the sexes. In the 39 individuals who had unilateral three-rooted mandibular first molars, 7 (17.9%) were on the left side and 32 (82.1%) were on the right side, indicating a statistically significant predominance on the right side. Conclusions The incidence of three-rooted mandibular first molars in contemporary Japanese individuals was 25.9%, with no statistically significant sex difference, but in the case of unilateral three-rooted teeth, with a statistically significant predominance on the right side. Our study found that Japanese and northeastern Asians have a high incidence of the three-rooted mandibular first molars among individuals of Mongolian origin; this finding may be a useful screening tool for identification of unknown individuals.

In this paper, we report a fatal case of multiple intracerebral hemorrhages associated with lipohyalinosis. A woman in her sixties was admitted to the hospital, where she was subsequently diagnosed with intracerebral hemorrhage. While hospitalized, she suffered several intracerebral hemorrhages and died 67 days after admission. During her hospital stay, she was also diagnosed with cerebral amyloid angiopathy (CAA). The results of autopsy and postmortem computed tomography indicated that the cause of death was a fourth hemorrhage from the cortex to the subcortex in the left parietal lobe. Microscopy further revealed eosinophilic and structureless material in the vessel walls of the brain surface. In addition, these blood vessel walls did not stain by Congo Red stain or immunohistochemical staining using the anti-amyloid β-protein antibody, but they were stained blue with the azocarmine stain. Thus, the decedent was considered to be in a state of lipohyalinosis. To the best of our knowledge, this is the first case report of a death after multiple intracerebral hemorrhages as a result of lipohyalinosis. Based on these findings, the possibility cannot be excluded that there may be a case that has been clinically diagnosed as CAA despite the true cause of multiple intracerebral hemorrhages being small vessel disease such as lipohyalinosis. Therefore, it is important to conduct brain tissue examinations to make a correct diagnosis.

To elucidate postmortem computed tomography (PMCT) and postmortem magnetic resonance (PMMR) imaging findings suggesting massive fat embolism.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.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.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.

Long QT syndrome (LQTS), a congenital genetic disorder, can cause torsades de pointes (TdP), and lethal cardiac arrhythmia may result from ingestion of cardiotoxic drugs. Methamphetamine (MP) and new psychoactive substances (NPSs) can trigger TdP due to QT prolongation, leading to sudden death. We therefore analysed variations in the LQTS-associated genes KCNQ1 (LQT1) and KCNH2 (LQT2) using cardiac blood and myocardial tissue from subjects having died suddenly during MP or NPS use to investigate the relationship between congenital genetic abnormalities and sudden death during illegal drug use. We amplified and sequenced all exons of these genes using samples from 20 subjects, half of whom had died taking MP and half after using NPSs. G643S, a KCNQ1 missense polymorphism, was significantly more common among sudden deaths involving NPSs (6 subjects) than those involving MP (1 subject) and healthy Japanese subjects (P = 0.001). Notably, synthetic cathinones were detected in 2 of 3 cases involving G643S carriers. Previous functional analyses have indicated that the G643S polymorphism in the KCNQ1 potassium channel gene causes mild IKs channel dysfunction. Our data suggest that use of NPSs, particularly synthetic cathinones, is associated with elevated risk of serious cardiac arrhythmia and sudden death for subjects carrying KCNQ1 G643S.