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.

Biomarker profiling of postmortem blood for diabetes mellitus and discussion of possible applications of metabolomics for forensic casework.

Acute metabolic disorders of diabetes mellitus (DM), such as diabetic ketoacidosis, hyperosmolar hyperglycemic state, and hypoglycemia, are life-threatening and difficult to diagnose postmortem owing to lack of macroscopic and microscopic findings, especially when the medical history of the patient is not available before autopsy. Although various biochemical tests, including ketone bodies and hemoglobin A1c, have been used to diagnose diabetes in the postmortem setting, each marker has some limitations. Consequently, it would be helpful in forensic practice to find new biomarkers reflecting the decedent's history of DM irrespective of whether the DM was being treated. Metabolomics enables the non-targeting analysis of biomarkers, and metabolomics was performed on postmortem blood from decedents with and without a DM history to determine whether a marker reflecting DM could be identified. The statistical analysis, including primary component analysis, presented a potent set of metabolites that could be used for the forensic diagnosis of DM. Qualitative analysis revealed significantly lower sphingomyelin and plasmalogen lipid levels and higher lysophospholipid levels in the DM group. Meanwhile, some discrepancies in the levels of some classes of phospholipids were noted between samples from living and deceased persons. This suggests that further metabolomics using postmortem samples rather than living persons' samples is required to identify markers that can be used for forensic diagnosis.

Age estimation by evaluation of osteophytes in thoracic and lumbar vertebrae using postmortem CT images in a modern Japanese population.

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.

Trends and Characteristics in Health Care-related Deaths Investigated through Medico-legal Autopsies after System Changes in Japan.

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.

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.

Fatal hemorrhage complicated with methamphetamine poisoning and its post-mortem CT features.

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.

Postmortem angiography revealing traumatic rupture of the intracranial internal carotid artery.

Diagnosis of traumatic subarachnoid hemorrhage (SAH), although relatively rare, is important in forensic medicine. It is mostly associated with rupture of the vertebrobasilar artery. Traumatic aneurysm of the intracranial part of the internal carotid artery (ICA) is also rare but has been reported in several studies. It is thought that the intracranial ICA is injured by blunt force to the head, neck, and chest. However, traumatic SAH with fatal acute course resulting from rupture of the ICA is especially uncommon: only two fatal cases without an associated aneurysm have been reported in the English-language literature. Although detecting the arterial lesion is required to make a precise diagnosis, this is sometimes impossible by macroscopic examination at autopsy or by investigation after formalin-fixation according to the position of the lesion. We report a rare case of fatal traumatic SAH associated with intracranial ICA rupture. Postmortem computed tomography angiography was useful to confirm the lesion.

Determination of sex on the basis of hyoid bone measurements in a Japanese population using multidetector computed tomography.

Sex determination for unidentified human remains is important because the determined sex is usually used as a foundation for the estimation of other biological profiles. This study investigated the sexual dimorphism of the adult hyoid bone and developed discriminant equations to accurately estimate sex in a contemporary Japanese population using measurements on two-dimensional computed tomography (CT) images. The study sample comprised 280 cadavers (140 males, 140 females) of known age and sex that underwent postmortem CT and subsequent forensic autopsy. In accordance with previous studies, seven measurements were performed using two-dimensional CT reconstructed images. Most measurements of males were significantly greater than those of females. The perpendicular length from the most anterior edge of the hyoid body to the line that connects the most distal points of the greater horns of the hyoid and the linear distance between the most lateral edges of the hyoid body most significantly contributed to sex determination. This study demonstrated that discriminant functions on the basis of three measurements provided higher rates of accurate sex classification (93.3-94.6%) than univariate functions. Therefore, the hyoid bone is highly sexually dimorphic in the contemporary Japanese population and may be useful in forensic contexts for sex determination with a high level of accuracy.

Stature estimation based on femoral measurements in the modern Japanese population: a cadaveric study using multidetector computed tomography.

We aimed to reproduce the anthropometrical measurement of femoral dimensions using multi-planar reconstruction computed tomography (CT), assess the correlation between stature and femoral measurements obtained by this approach, and establish a regression equation for estimating stature in the modern Japanese population. We used data regarding 224 cadavers (116 males, 108 females) that were subjected to postmortem CT and subsequent forensic autopsy at our department between October 2009 and July 2016. To simulate the placement of the femur on the osteometric board using reconstructed CT images, we defined a virtual horizontal plane (VHP) based on the three most dorsal points of the femur (lateral condyle, medial condyle, and greater trochanter). Five femoral measurements including the maximum femoral length (MFL) were obtained. The correlations between stature and each femoral measurement were expressed in terms of the coefficient of determination (R2). On regression analysis, MFL provided the lowest value for the standard error of the estimation (SEE); the SEE values in all subjects, males, and females, respectively, were 3.783 cm (R2 = 0.832), 3.850 cm (R2 = 0.653), and 3.340 cm (R2 = 0.760) for MFL on the left side and 3.747 cm (R2 = 0.835), 3.847 cm (R2 = 0.650), and 3.290 cm (R2 = 0.687) for MFL on the right side. Multiple regression equations using MFL and femoral epicondylar breadth were slightly superior to simple regression equations in males and in all subjects (SEE = 3.44-3.55 cm), whereas no effective equation could be obtained in females. To our knowledge, this is the first multiple regression equation for stature estimation using only femoral measurements.

Myoglobinemia markers with potential applications in forensic sample analysis: lipid markers in myoglobinemia for postmortem blood.

The crush syndrome, in which rhabdomyolysis and trauma occur as a result of heat stroke and drug intoxication, can lead to myoglobinemia. This condition can be diagnosed by measuring myoglobin (Mb) levels in blood and urine. However, postmortem Mb levels are unreliable indicators, since blood Mb concentration drastically increases within a very short time after death and urine cannot always be obtained at dissection; this makes it difficult to diagnose myoglobinemia in a corpse. To address this issue, in this study, we used a lipidomics approach to identify markers that can be used to detect myoglobinemia in postmortem blood samples. We found that increases in levels of fatty acid oxides such as stearic, oleic, linoleic, and arachidonic acid and decreases in levels of plasmalogens and phosphatidylethanolamine in the blood were associated with high Mb level. These results demonstrate that postmortem samples are amenable to lipidomics analysis and provide a set of markers other than Mb that can be used for postmortem diagnosis of myoglobinemia.

Frequency and influencing factors of cardiopulmonary resuscitation-related injuries during implementation of the American Heart Association 2010 Guidelines: a retrospective study based on autopsy and postmortem computed tomography.

AIM: To determine the frequency of cardiopulmonary resuscitation (CPR)-related injuries and factors involved in their occurrence, data based on forensic autopsy and postmortem computed tomography (PMCT) during implementation of the 2010 American Heart Association Guidelines for CPR were studied. METHODS: We retrospectively evaluated data on adult patients with non-traumatic deaths who had undergone manual CPR and autopsy from January 2012 to December 2014. CPR-related injuries were analyzed on autopsy records and PMCT images and compared with results of previous studies. RESULTS: In total, 180 consecutive cases were analyzed. Rib fractures and sternal fractures were most frequent (overall frequency, 66.1 and 52.8%, respectively), followed by heart injuries (12.8%) and abdominal visceral injuries (2.2%). Urgently life-threatening injuries were rare (2.8%). Older age was an independent risk factor for rib fracture [adjusted odds ratio (AOR), 1.06; 95% confidence interval (CI), 1.04-1.08; p < 0.001], ≥ 3 rib fractures (AOR, 1.06; 95% CI, 1.02-1.09; p = 0.002), and sternal fracture (AOR, 1.03; 95% CI, 1.01-1.05; p < 0.001). Female sex was significantly associated with sternal fracture (AOR, 2.08; 95% CI, 1.02-4.25; p = 0.04). Chest compression only by laypersons was inversely associated with rib and sternal fractures. Body mass index and in-hospital cardiac arrest were not significantly associated with any complications. The frequency of thoracic skeletal injuries was similar to that in recent autopsy-based studies. CONCLUSIONS: Implementation of the 2010 Guidelines had little impact on the frequency of CPR-related thoracic skeletal injuries or urgently life-threatening complications. Older age was the only independent factor related to thoracic skeletal injuries.

A case of acute subdural hematoma due to ruptured aneurysm detected by postmortem angiography.

Acute subdural hematoma (ASDH) is mostly caused by head trauma, but intrinsic causes also exist such as aneurysm rupture. We describe here a case involving a man in his 70s who was found lying on the bedroom floor by his family. CT performed at the hospital showed ASDH and a forensic autopsy was requested. Postmortem cerebral angiography showed dilatation of the bifurcation of the middle cerebral artery, which coincided with the dilated part of the Sylvian fissure. Extravasation of contrast medium into the subdural hematoma from this site was suggestive of a ruptured aneurysm. Autopsy revealed a fleshy hematoma (total weight 110 g) in the right subdural space and findings of brain herniation. As indicated on angiography, a ruptured saccular aneurysm was confirmed at the bifurcation of the middle cerebral artery. Obvious injuries to the head or face could not be detected on either external or internal examination, and intrinsic ASDH due to a ruptured middle cerebral artery aneurysm was determined as the cause of death. One of the key points of forensic diagnosis is the strict differentiation between intrinsic and extrinsic onset for conditions leading to death. Although most subdural hematomas (SDH) are caused by extrinsic factors, forensic pathologists should consider the possibility of intrinsic SDH. In addition, postmortem angiography can be useful for identifying vascular lesions in such cases.

Fatal right coronary artery rupture following blunt chest trauma: detection by postmortem selective coronary angiography.

Coronary artery injury is a rare complication following blunt chest trauma (BCT), and can be fatal. Here we report findings on postmortem selective coronary angiography of right coronary artery rupture after an assault involving blunt trauma to the chest. A woman in her 60s died after her son stomped on her chest. There were no appreciable signs of injury on external examination, and cause of death could not be determined by postmortem computed tomography (PMCT). Internal findings indicated that an external force had been applied to the anterior chest, as evidenced by subcutaneous hemorrhage and pericardial and cardiac contusions. Postmortem coronary angiography revealed irregularity of the intima and of the fat tissue surrounding the proximal part of the right coronary artery associated with a local filling defect. Histopathological examination suggested coronary rupture with dissection of the tunica media and compression of the lumen cavity. The key points in the present case are that no fatal injuries could be determined on external examination, and the heart and coronary artery injuries were not evident on PMCT. Criminality might be overlooked in such cases, as external investigation at the crime scene would be inadequate and could result in a facile diagnosis of cause of death. This is the first report of coronary artery rupture with dissection that was detected by CT coronary angiography, and provides helpful findings for reaching an appropriate decision both forensically and clinically.

Evaluations of lipid peroxidation and inflammation in short-term glycerol-induced acute kidney injury in rats.

Rhabdomyolysis is characterised by acute kidney injury (AKI) resulting from skeletal muscle injury. Lipid peroxidation-mediated oxidant injury and pro-inflammatory cytokine-mediated inflammatory response play critical roles in the pathogenesis of rhabdomyolysis-induced AKI. The present study aimed to investigate the short-term effects of both lipid peroxidation and inflammatory responses on rhabdomyolysis-induced AKI in a rat model of glycerol-induced rhabdomyolysis. Rhabdomyolysis was induced by the intramuscular injection of 50% glycerol in saline (10 mL/kg) into the hind limbs of rats. Rats were killed 1 or 3 hours after glycerol injection. Time-dependent increases in serum biochemical parameters, including blood urea nitrogen, creatinine, lactate dehydrogenase and creatine phosphokinase levels, were observed 1 hour after glycerol injection. In kidneys, glycerol injection resulted in histopathological changes such as renal tubular injury and renal tubular myoglobin deposition. Levels of Nε-(hexanoyl)lysine-modified, 4-hydroxy-2-nonenal-modified, and nitrotyrosine-modified proteins in rat kidneys were unaltered at 1 hour after glycerol injection, but increased significantly at 3 hours. Increases in renal nitric oxide production and the expression levels of inducible nitric oxide synthase, interleukin-6 and tumour necrosis factor-α in the renal parenchyma were observed at 1 hour after glycerol injection and plateaued at 3 hours. Our findings suggest that the pro-inflammatory cytokine-mediated inflammatory response may cause rhabdomyolysis-induced AKI very shortly after glycerol injection, and lipid peroxidation-mediated oxidant injury may promote the development of these pathophysiological processes.

Stature estimation in Japanese cadavers based on scapular measurements using multidetector computed tomography.

This study assessed the correlation between stature and scapular measurements in a Japanese population, using three-dimensional (3D) computed tomographic (CT) images, and derived regression equations for predicting stature. A total of 194 cadavers (100 males 94 females) underwent postmortem CT (PMCT) and subsequent forensic autopsy in our department between May 2011 and April 2014. Left and right longitudinal scapular lengths (LLSL and RLSL, respectively) and left and right transverse scapular lengths (LTSL and RTSL, respectively) were measured on 3D CT reconstructed images that extracted only scapular data. The correlation between stature and each of the scapular measurements were analyzed using Pearson product-moment correlation coefficients. The four variables correlated significantly with stature, regardless of sex. The LLSL measurement had the lowest standard error of estimation value among all subjects (4.22 cm) and among all females (4.37 cm), whereas the RLSL measurement had the lowest standard error of estimation value among all males (3.75 cm). The results of this study indicate that scapular measurements may be useful for the forensic estimation of the stature of Japanese individuals, particularly in cases where better predictors, such as long bone lengths, are unavailable.

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.

Gray-white matter contrast reversal on T1-weighted spin-echo in postmortem brain.

PURPOSE: The image contrast of postmortem magnetic resonance imaging (MRI) may differ from that of antemortem MRI because of circulator arrest, changes in postmortem tissue, and low-body-temperature scanning conditions. In fact, we have found that the signal intensity of white matter (WM) on T1-weighted spin-echo (T1WSE) images of the postmortem brain was lower than that of gray matter (GM), which resulted in image contrast reversal between GM and WM relative to the living brain. However, the reason for this phenomenon is unclear. Therefore, the aim of this study is to clarify the reason why image contrast reversal occurs between GM and WM of the postmortem brain. MATERIALS AND METHODS: Twenty-three corpses were included in the study (mean age, 60.6 years; range: 19-60 years; mean rectal temperature at scan, 6.9℃; range: 4-11℃). On a 1.5 T MRI system, postmortem T1W-SE MRI of the brain was conducted in the 23 corpses prior to medico-legal autopsy. Next, T1 and T2 of the GM and WM at the level of the basal ganglia were determined in the same participants using inversion recovery and multiple SE sequences, respectively. The proton density (PD) was also calculated from the T1 and T2 images (in the same slice). RESULTS: T1W-SE image contrast between the GM and WM of all postmortem brains was inverted relative to the living brain. T1 (579 ms in GM and 307 ms in WM) and PD (64 in GM and 44 in WM) of the postmortem brain decreased compared with the living brain. While T1 of WM/GM remained below 1 even postmortem, the PD of WM/GM decreased. T2 (110 ms in GM and 98 ms in WM) of the postmortem brain did not differ from the living brain. CONCLUSION: The decrease in PD of WM/GM in the postmortem brain may be the major driver of contrast reversal between the GM and WM relative to the living brain.

Identification of postmortem paliperidone metabolite in human blood by LC-Q-Orbitrap-MS.

Paliperidone is a widely used antipsychotic agent detected in many fatal intoxications and suicide cases. In forensic toxicology, the accurate determination of blood paliperidone concentrations is required to prove death by paliperidone poisoning. However, the lethal concentration of paliperidone in blood at autopsy differs from that at the time of death. In this study, we found that paliperidone was decomposed by hemoglobin (Hb) through the Fenton reaction in a temperature-dependent fashion. The mechanism underlying paliperidone decomposition involves the cleavage of its C-N bond linker moiety. The mass spectra obtained by liquid chromatography-quadrupole orbitrap mass spectrometry revealed the formation of 6-fluoro-3-(4-piperidinyl)benzisoxazole (PM1) in Hb/H2O2 solution incubated with paliperidone, as well as in the blood of individuals who died from intentional ingestion of paliperidone. These results suggest that PM1 is the only metabolite produced from paliperidone as a result of temperature-dependent, postmortem changes induced by Hb via the Fenton reaction and may be useful as a biomarker to correct for the concentration of paliperidone in blood at the time of death in clinical cases.

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.