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日本においては，法医学をはじめ，法と医のインターフェース的な役割を担う領域が軽視されてきた．医療関連死にかかわるさまざまな問題も，そうしたことと無関係とはいえないだろう．医師が司法の介入でストレスを感じる場合，その対策としては，司法を排除する方法以外にも，司法の暴走を抑止し，適正な司法の実現に協力するという方策もありうる．現在，医師だけでなく，司法もミスを犯す存在であることは周知の事実となりつつあり，さまざまな司法制度改革も始まっている．そのなかで，医師，法曹ともに，法と医のインターフェース的役割の在り方を見つめなおし，適正な司法を実現するためには，医学鑑定がどうあるべきかなどを検討してもよいだろう．

We consecutively observed lipid peroxidation and cell membrane damage under the condition of hypoxia-reoxygenation (H/R) in cells and analyzed their mechanisms by using electron transport inhibitors and an antioxidant. In H/R experiments, lipid peroxidation and cell membrane damage were observed during the hypoxia phase. In the reoxygenation phase, lipid peroxidation stopped, while cell membrane damage did not. An antioxidant, n-acetylcystein (NAC), and potassium cyanide (KCN) inhibited lipid peroxidation and cell membrane damage, while rotenone did not inhibit either of them. Although antimycin A did not inhibit lipid peroxidation, it inhibited cell membrane damage during the hypoxia phase but not during the reoxygenation phase. These results suggested that lipid peroxidation can affect cell membrane damage as a trigger during the hypoxia phase and the generation of oxidative stress can vary depending on the inhibition locations in the electron transport system.

We discuss the usefulness of postmortem computed tomography (PMCT) by reviewing cases of cervical spine injury. A merit of PMCT is that it can identify injury that cannot be found on autopsy; however, peculiar defects of it may exist. While PMCT can identify bone fractures, it cannot indicate whether the injury was inflicted while the deceased was still alive or not because of its inability to clearly image bleeding around the fracture. Furthermore, CT often misses some types of cervical spine injuries, such as laceration of an intervertebral disk and incomplete fracture of the cervical spine. On the other hand, cervical spine injury on CT images occasionally has an appearance similar to subarachnoid hemorrhage due to rupture of the cerebral artery, indicating that cervical spine injury can be misdiagnosed as a disease by PMCT. When PMCT is used for screening trauma, caution must be observed regarding its limitations. If the possibility of trauma of the neck or head is not completely ruled out from the personal history of the victim, autopsy is strongly recommended, even when PMCT findings indicate that the cause of death may be due to disease, such as subarachnoid hemorrhage.

Purpose The purpose of this study is to assess the applicability of facial soft tissue measurements using 3-dimensional reconstructed MDCT and to compare the results with previously reported Japanese data. Methods and materials This study included 50 Japanese cadavers who underwent postmortem whole body 16-detector CT within 0–3 days after death. Using 3-D workstation, 10 anthropological measuring points were located and the soft tissue thicknesses were measured. The differences between the acquired thickness and the previously reported Japanese average thickness were statistically analyzed. Results All measuring points were readily determined on the 3-dimensional reconstructed images. Previous Japanese cadaveric data was thinner than our data in almost all sites except for 1 measuring point (mid-philtrum). Previous living Japanese data was thicker than ours except for 1 measuring point (end of nasal). Conclusion Facial soft tissue thickness was easily assessed using 3-D reconstructed MDCT images. At many measuring points, our results were thicker than previously reported cadaveric data and were thinner than the data from live persons.

17β-Estradiol (E2–17β), the female hormone with the strongest estrogenic activity, is thought to be present in high quantities in vaginal secretions because it is synthesized mainly in the ovaries and placenta. To clarify whether the detection of E2–17β is useful for identifying vaginal secretions among other body fluid stains, we quantified the amount of E2–17β in extracts from body fluid stains of approximately 4 cm in diameter using gas chromatography-mass spectrometry (GC-MS). The results showed that E2–17β was strongly detected in all of 6 extracts from vaginal stains (81–1,811 ng/mL), 1 of 10 extracts from semen stains (31 ng/mL), and 1 of 9 extracts from female saliva stains (37 ng/mL). All other samples tested were below the limit of quantification. In addition, E2–17β did not show significant degradation in stains kept for 2 days at room temperature. Then, the usefulness of a simple and rapid commercial enzyme-linked immunosorbent assay (ELISA) kit using E2–17β monoclonal antibody was exa...

We report a fatal case of hypovolemic shock caused by uncontrollable hemorrhaging after emergency cesarean section. In this patient, the incision in the uterus was located only 1 cm from the cervical os. We suspect that this close incision was the cause of the damage to the uterine venous plexus and the bleeding. We discuss the cause of death and offer advice on performing autopsies in patients who have died of bleeding after cesarean section.

Nitric oxide (NO) has been implicated as a cardioprotective agent during ischemia/reperfusion (I/R), but the mechanism of protection is unknown. Oxidant stress contributes to cell death in I/R, so we tested whether NO protects by attenuating oxidant stress. Cardiomyocytes and murine embryonic fibroblasts were administered NO (10-1200 nM) during simulated ischemia, and cell death was assessed during reperfusion without NO. In each case, NO abrogated cell death during reperfusion. Cells overexpressing endothelial NO synthase (NOS) exhibited a similar protection, which was abolished by the NOS inhibitor N(omega)-nitro-l-arginine methyl ester. Protection was not mediated by guanylate cyclase or the mitochondrial K(ATP) channel, as inhibitors of these systems failed to abolish protection. NO did not prevent decreases in mitochondrial potential, but cells protected with NO demonstrated recovery of potential at reperfusion. Measurements using C11-BODIPY reveal that NO attenuates lipid peroxidation during ischemia and reperfusion. Measurements of oxidant stress using the ratiometric redox sensor HSP-FRET demonstrate that NO attenuates protein oxidation during ischemia. These findings reveal that physiological levels of NO during ischemia can attenuate oxidant stress both during ischemia and during reperfusion. This response is associated with a remarkable attenuation of cell death, suggesting that ischemic cell death may be a regulated event.

The aim of this study was to assess the correlation between stature and cranial measurements in a contemporary Japanese population, using three-dimensional (3D) computed tomographic (CT) images. A total of 228 cadavers (123 males, 105 females) underwent postmortem CT scanning and subsequent forensic autopsy between May 2011 and April 2015. Five cranial measurements were taken from 3D CT reconstructed images that extracted only cranial data. The correlations between stature and each of the cranial measurements were assessed with Pearson product-moment correlation coefficients. Simple and multiple regression analyses showed significant correlations between stature and cranial measurements. In conclusion, cranial measurements obtained from 3D CT images may be useful for forensic estimation of the stature of Japanese individuals, particularly in cases where better predictors, such as long bones, are not available.

A man with tattoos was found dead in the sea. Although the body was highly decomposed, his fingerprints identified him as a man in his 40s known to have a history of amphetamine abuse. A week prior to being found, he had stated that he wanted to commit suicide after having made a mistake at work. Video images from security cameras suggested that he drove to the sea alone. Whole-body postmortem computed tomography (PMCT) scans followed by a forensic autopsy were performed. The CT protocol used the following scan parameters: 120 kV tube voltage, 200 mA tube current, 1 rotation per second, 1.25 mm collimation, and a pitch of 0.93. PMCT showed 14 small, high-density particles that appeared to have slight metallic artifacts: 11 particles in the right lobe of the lung and 3 in the wall of the right ventricle (Fig. 1). The particles were 1–2 mm in diameter. The CT values of the particles were 179–1065 Hounsfield units (HU) (mean 517 HU, median 545 HU). Autopsy revealed fluid with high electrolyte concentrations in both pleural cavities. Diatoms were found in both lungs but not in the kidneys. Toxicologic screening tests of blood, muscle, and urine samples, using liquid chromatography-tandem mass spectrometry, were all negative. To identify the high-density particles revealed by PMCT, small specimens that included some of the particles each were extracted, using CT guidance, from formalin-fixed right lung tissue and from right ventricular wall tissue. X-ray fluorescence (XRF) analysis with an X-ray analytic microscope (XGT-5200; Horiba, Kyoto, Japan) was performed to determine the composition of the particles. This machine has a thin (10 lm at its thinnest) glass tube that can narrow the X-ray beam. The beam is focused on a sideby-side basis on the tissue specimen; enabling element mapping by visualizing XRF results point by point with a resolution of 10–100 lm. The XGT-5200 protocol included the following parameters: 100 lm glass tube diameter, 50 kV tube voltage, and 1000 s/frame acquisition time. The XGT-5200 results after scanning one frame are shown in Fig. 2a–d (lung) and Fig. 3a–d (heart). Both analyses detected fluorescence consistent with that of mercury in the same positions where there was high X-ray attenuation on CT findings (Figs. 2e, 3e). Point analysis showed a high-energy spectrum of mercury, whereas there was no mercury spectrum at distant points. None of the spectra of other metals, including those for calcium, iron, and sodium, were significantly changed between these points. Therefore, we concluded that the X-ray absorption & Yohsuke Makino ymakino-tky@umin.ac.jp

Pediculus humanus humanus (known as body lice) are commonly found in the folds of clothes, and can cause skin disorders when they feed on human blood, resulting in an itching sensation. Body lice are known as vectors of infectious diseases, including typhus, recurrent fever, and trench fever. An infestation with blood-sucking body lice induces severe cutaneous pruritus, and this skin disorder is known as "vagabond's disease." A body lice infestation is sometimes complicated with iron deficiency anemia. In the present case, a man in his late 70s died of lethal hypothermia in the outdoors during the winter season. The case history and autopsy findings revealed that the cause of the lethal hypothermia was iron deficiency anemia, which was associated with a prolonged infestation of blood-sucking body lice. Also, he had vagabond's disease because the skin on his body was abnormal and highly pigmented. This is an unusual autopsy case since the body lice contributed to the cause of the death.

Sex estimation of decomposed or skeletal remains is clearly important in forensic contexts. Recently, contemporary population-specific data has been obtained using multidetector computed tomography (MDCT) scanning. The main purpose of this study was to investigate skeletal pelvic dimorphism in a contemporary Japanese forensic sample and to quantify the accuracy of sex estimation using various pelvic measurements obtained from three-dimensional (3D) CT images. This study used a total of 208 cadavers (104 males, 104 females) of which postmortem CT scanning and subsequent forensic autopsy were conducted between December 2011 and August 2014. Eleven measurements of each pelvis were obtained from 3D CT reconstructed images that extracted only bone data. The measurements were analyzed using descriptive statistics and discriminant function analyses. All except one measurement were dimorphic in terms of sex differences. Univariate discriminant function analyses using these measurements provided sex classification accuracy rates of 62.0-98.1%. The subpubic angle was found to contribute most significantly to accurate sex estimation. Multivariate discriminant functions yielded sex prediction accuracy rates of 63.9-98.1%. In conclusion, the pelvic measurements obtained from 3D CT images of a contemporary Japanese population successfully demonstrated sexual dimorphism and may be useful for the estimation of skeletal sex in the field of forensic anthropology.

Forensic toxicology is aimed at identifying the relationship between drugs or poison and the cause of death or crime. In the authors' toxicology laboratory at Chiba University, the authors analyze almost every body for drugs and poisons. A simple inspection kit was used in an attempt to ascertain drug abuse. A mass spectrometer is used to perform highly accurate screening. When a poison is detected, quantitative analyses are required. A recent topic of interest is new psychoactive substances (NPS). Although NPS-related deaths may be decreasing, use of NPS as a cause of death is difficult to ascertain. Forensic institutes have recently begun to perform drug and poison tests on corpses. However, this approach presents several problems, as are discussed here. The hope is that highly accurate analyses of drugs and poisons will be performed throughout the country.

The aim of this study was to assess the mechanical properties and thickness of adult frontal and parietal bones. The heads of 114 Japanese cadavers (78 male cadavers and 36 female cadavers) of known age and sex were used. A total of 912 cranial samples, 8 from each skull, were collected. Samples were imaged using multidetector computed tomography to measure sample thickness. The fracture load of each sample was measured using a bending test with calculation of flexural strength. Statistical analyses demonstrated no significant bilateral difference in either the mechanical properties or thickness of frontal or parietal bones. The mechanical properties and thicknesses of frontal bones were significantly greater than those of parietal bones regardless of sex. Therefore, the skull may have a great ability to resist frontal impacts compared with parietal impacts. In female samples, parietal bones were found to have a more uniform structure when compared with male samples. Male parietal bones were found to be thicker at medial sites than at lateral sites. This study also revealed parietal bones at lateral sites in female samples were thicker than in male samples. No strong association was observed between age and flexural strength of frontal or parietal bones. However, the fracture load was negatively correlated with age most likely due to the reduction of thickness.

We present an autopsy case of sudden death due to coronary artery dissection associated with fibromuscular dysplasia (FMD) in a young female patient. Postmortem selective coronary artery computed tomography (CT) angiography revealed dissections of the left anterior descending and left circumflex arteries. These findings were confirmed by subsequent autopsy. Histopathological examination revealed coronary artery FMD, which is considered a risk factor for dissection. To the best of our knowledge, this is the first postmortem radiology-pathology correlation of coronary artery dissection associated with FMD.

Donepezil (DPZ) is an acetylcholine-esterase inhibitor currently used as the frontline drug to treat Alzheimer's disease. The aim of this study was to investigate the possibility of postmortem redistribution (PMR) of DPZ, which could complicate the determination of cause of death in medico-legal cases. Additionally, metabolic enzyme DNA polymorphism, drug-drug interaction and the presence of lesions in metabolic and egestion organs were examined to eliminate the possibility of a high antemortem DPZ concentration. Subsequently, the average DPZ concentration of four sites of peripheral blood (right and left femoral artery and vein) was compared with central blood and liver DPZ concentrations in seven postmortem cases. DPZ concentrations ranged from 0.02 to 0.45μg/mL in the peripheral blood, 0.09-0.4μg/mL in central blood, and from 1.2 to 6.7μg/kg in the liver. In most specimens, the concentrations were higher than the therapeutic range (approximately 0.030-0.075μg/mL). DPZ central blood to peripheral blood (C/P) ratios averaged 1.73±1.02 (±standard deviation) while liver to peripheral blood (L/P) ratios were higher and averaged 17.5±7.25. It is documented that a C/P ratio of less than (or about) 1.0 and an L/P ratio less than 5 are not indicative of PMR, whereas a C/P ratio exceeding 2 and L/P ratio exceeding 20 highlight a propensity for significant PMR. Our data suggest that DPZ exhibits a moderate degree of PMR. Additionally, a lowered pH was found in all blood specimens (

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.

We examined the usefulness of the dura mater in identifying human remains. Dura mater was collected from 50 cadavers, including drowned, charred, and mummified remains. The STR genotype using the AmpFlSTR® Identifiler™ Kit could be typed at 15 STR and amelogenin loci in 30 samples of 33 cases. Furthermore, the ABO genotype and amelogenin using gel-based methods could be typed in 44 samples of 50 cases. In cases with successful typing of STR, ABO-DNA, and amelogenin, the longest time after death was from 12 to 26 days in a drowned body. The minimum quantity of dura mater required for DNA extraction was about 2.5 mg, dried and fixed by ethanol, in a cadaver 15 h after death. The state of the DNA from the dura mater from the calvaria may be better than that from the basis cranii interna. We found that DNA from dura mater is one of the most useful samples for forensic identification.

A genotype of JC virus (JCV) DNA extracted from human urine can serve not only as a means of elucidating human migration patterns, but also of tracing the origin of unidentified cadavers. If a JCV genotype is determined using a small amount of urine or urine stain from a given person, it may be useful in identifying the geographic area in which that person was born. In the present study, we amplified a 610-bp intergenic region (IG region) of JCV DNA extracted from 200 μl of each of 82 urine samples collected in Japan. JCV DNA was detected in 45 of 82 urine samples (54.9%) and genotypes were also determined by restriction fragment length polymorphism analysis (MY genotype: 13; X (CY) genotype: 25; unclear: 7). Additionally, 16 urine stains which had been prepared 3 months earlier using 100 μl of JCV-positive urine were examined. JCV DNA was detected in 11 of 16 urine stains (68.8%), and in this case also, it was possible to determine the genotype by the phylogenetic analysis after DNA sequencing. The present study indicates that JCV genotyping can be successfully conducted using only 100 μl urine stain and even 3 months after the urine stain was made. Thus, JCV genotyping shows promise as a new analytical method for narrowing down the native place of unidentified persons or cadavers from urine stains left at the scene.

We studied JC virus (JCV) DNA sequence diversity among kidneys derived from cadavers with various causes of death. The 610-bp JCV DNA sequences we evaluated were identical not only among specimens derived from the same kidney but also among those derived from both kidneys of the same cadaver. Because the left and right kidneys are anatomically independent, our findings suggest that the viremia that has been proposed to occur after primary infection distributes the same JCV strain to both kidneys.

To preliminarily evaluate the applicability of bacterial DNA as a marker for the forensic identification of vaginal fluid, we developed and performed PCR-based detection of 16S ribosomal RNA genes of Lactobacillus spp. dominating the vagina and of bacterial vaginosis-related bacteria from DNA extracted from body fluids and stains. As a result, 16S ribosomal RNA genes of Lactobacillus crispatus, Lactobacillus jensenii and Atopobium vaginae were specifically detected in vaginal fluid and female urine samples. Bacterial genes detected in female urine might have originated from contaminated vaginal fluid. In addition, those of Lactobacillus iners, Lactobacillus gasseri and Gardnerella vaginalis were also detected in non-vaginal body fluids such as semen. Because bacterial genes were successfully amplified in DNA samples extracted by using the general procedure for animal tissues without any optional treatments, DNA samples prepared for the identification of vaginal fluid can also be used for personal identification. In conclusion, 16S ribosomal RNA genes of L. crispatus, L. jensenii and A. vaginae could be effective markers for forensic identification of vaginal fluid.

There has been no reliable means of tracing the origins of unidentified cadavers but the recent finding that JC virus (JCV) can serve as a means of elucidating human migrations suggested that this virus may also be useful to trace the origins of unidentified cadavers. DNA samples extracted from renal tissue and urine were used as the template for PCR amplification of a 610 bp region (IG region) of the viral genome. We detected JCV DNA in 45% of the renal samples and in 33% of the urine samples and was detectable even 10 days after death. The sequences of the amplified IG regions could be used to determine the genotypes. We conclude that the JC virus genotype is a new marker useful for tracing the origins of unidentified cadavers.

Gamma-hydroxybutyric (GHB) acid, which is becoming popular as a drug of abuse, was shown by means of gas chromatography-mass spectrometry (GC-MS) to increase in mouse liver with time after death. The amount detected was 0.8 +/- 1.0 microg/g at 3 h after death, 4.7 +/- 1.5 microg/g at 24 h, and 8.8 +/- 0.8 microg/g at 72 h. Furthermore, GHB was detected in samples from deceased persons, at concentrations of 2.6-12.0 microg/g in liver, 0.4-7.3 microg/ml in blood, and 0-2.6 microg/ml in urine, but was not detected in the blood and urine of living persons. Although 1,4-butanediol has been suggested to be a precursor of GHB produced after death, 1,4-butanediol was not detected in any of our samples. Additionally, succinate semialdehyde arising from gamma-aminobutyric acid (GABA) transamination to GHB was also barely detectable in any of our samples. This study supports previous reports that GHB is a product of postmortem decomposition. Production of GHB increases with time after death in postmortem liver; since we were unable to identify endogenous 1,4-butanediol and succinate semialdehyde in our samples, the pathway of GHB production after death remains unclear.

We applied the fusion function of dental analysis software to examine whether the anatomical relationships of tooth roots when compared between reconstructed post-mortem CT (PMCT) and dental radiographs can aid dental identification.One PMCT image taken from a cadaver (43-year-old male; Cadaver 1) was compared with 64 digital dental radiographs of the left and right upper and lower molars from Cadaver 1 and 30 other cadavers. Five corresponding anatomical reference points were marked on each image. After adjusting the angle and magnification using the fusion function, the automatically calculated error in pixels was determined five times for each of four sites on the images.Comparison of the PMCT image with the dental radiographs from the other cadavers revealed obvious discrepancies in the anatomical positioning of the teeth. When t-tests were applied to the data from any of the four sites, the error in pixels was found to be significantly smaller (p 0.001) between Cadaver 1 and the other cadaver images. The average error in pixels between the PMCT and dental radiographs was smaller in the lower jaw than in the upper jaw.This method uses corresponding reference points on two images to calculate the error between the regions that contain all points. This feature also makes it possible to compare images taken with different modalities. The demand for a dental identification method involving PMCT is likely to increase, and we expect that the accuracy of dental identification will improve by using radiological images.

The purpose of this study was to assess the incidence of spinal cord injuries without radiographic abnormalities (SCIWORA) on postmortem MDCT studies and to characterize this entity in detail.Data from consecutive autopsies from October 2009 through December 2012 were searched, yielding 30 cases of cervical spinal cord injuries. Two board-certified radiologists interpreted postmortem MDCT images acquired before autopsy. Cases with normal findings were defined as SCIWORA.Postmortem MDCT images in 6 of 30 cases (20%; 95% CI, 8-39%) met SCIWORA criteria. In all these cases, traumatic death was not apparent before CT, the highest injury level was below C3, and fractures were not identified at autopsy. All subjects had occult disk injuries and perivertebral hemorrhage. Potentially fatal CT-detectable injuries to other parts of the body were not seen in five of six SCIWORA cases.A substantial proportion of fatal spinal cord injuries are without radiographic abnormalities at CT. Forensic radiologists and other interpreters of postmortem MDCT images must be aware of this entity to avoid excluding spinal cord injuries solely on the basis of postmortem MDCT findings. Special care is necessary in cases of unknown circumstances of death without abnormal CT findings. MRI may be used for further examination in the future, but until then, MDCT cannot replace autopsy for detecting SCIWORA.

The purpose of this study was to assess correlations between measurements of the second cervical vertebra (C2) and stature using multidetector computed tomography (MDCT) images, and to develop regression equations for estimating stature in a Japanese population. Measurements were performed on 216 Japanese subjects (116 males and 100 females) who underwent postmortem CT between May 2011 and November 2013. Sagittal images through the center of the C2 were used for assessment. The length from the top of the dens to the anteroinferior point of the vertebral body (DA), the length from the anteroinferior point of the vertebral body to the posterior point of the spinous process (AS), and the length from the top of the dens to the posterior point of the spinous process (DS) were measured. The correlation between stature and each parameter (DA, AS, and DS) was assessed using Pearson product-moment correlation coefficients and regression analysis was performed for stature estimation. All measurements of the C2 were positively correlated with stature regardless of sex. The highest correlation was observed for the DA in all cases, and the lowest correlation was observed for AS in all cases. However, the standard errors of estimate were large. Thus, our study concludes that the size of the C2 as measured with MDCT images may be useful for stature estimation only when better predictors, such as long bones, are unavailable.

Stature estimation using a skeleton is important for the medicolegal investigation of unidentified human remains. The aims of this study were to identify a correlation between stature and measurements of the sternal medullary cavity using multidetector computed tomography (MDCT) and derive regression equations for stature estimation in the Japanese population. Measurements were conducted on 215 Japanese subjects (107 males, 108 females) who underwent postmortem computed tomography with subsequent forensic autopsy between May 2012 and January 2014. For assessment, MDCT cross-sections through the mid-point of the first costal facets were chosen. The length of a rising diagonal stroke from the bottom left to the top right of the sternal medullary cavity (RS) and the length of a falling diagonal stroke from top left to bottom right of the sternal medullary cavity (FS) were measured. Statistical analyses indicated that both RS and FS were positively correlated with stature regardless of sex. The correlations were stronger for males than for females. The correlation coefficients for RS were higher than those for FS, and standard errors of estimation calculated by regression analysis using RS were lower than those using FS regardless of sex. Measurement of the sternal medullary cavity using MDCT images may be a potentially useful tool for stature estimation, particularly in cases where better predictors such as long bones are not available.

There is a hypothesis suggesting that rigor mortis progresses more rapidly in small muscles than in large muscles. We measured rigor mortis as tension determined isometrically in rat musculus erector spinae that had been cut into muscle bundles of various volumes. The muscle volume did not influence either the progress or the resolution of rigor mortis, which contradicts the hypothesis. Differences in pre-rigor load on the muscles influenced the onset and resolution of rigor mortis in a few pairs of samples, but did not influence the time taken for rigor mortis to reach its full extent after death. Moreover, the progress of rigor mortis in this muscle was biphasic; this may reflect the early rigor of red muscle fibres and the late rigor of white muscle fibres.

A 25-year-old man was killed by his lover by an intravenous injection of insulin and then air. We had difficulty in determining whether insulin had really been injected, so we have 18 control cases and proved that the ratio of insulin to C-peptide in a corpse can be used as positive evidence for the insulin overdosage.

In this study, we investigated lipid peroxidation in rat heart mitochondria hydrolyzed by phospholipase A2 (PLA2) and lipid peroxidation in a mitochondrial-mimetic lipid peroxidation system, where phospholipids such as cardiolipin (CL) and cytochrome c (Cyt c) were first mixed together and then PLA2 and calcium chloride were added to the mixture (CL-Cyt c-PLA2 system). Production of hydroperoxy and hydroxy compounds of linoleic acid (LA) in the mixture was measured by high performance liquid chromatography. The ratio of the total amount of hydroperoxy and hydroxy compounds of LA to that of LA was calculated as an index for lipid peroxidation (1000 x mol/mol). The index for lipid peroxidation in the rat heart mitochondria hydrolyzed by PLA2 at the physiological pH of 7.4 was 22.8 +/- 2.2 (mean +/- SD, n = 4) and that at the acidic pH of 6.7 was 41.8 +/- 2.0. In the presence of the thiol (SH)-oxidizing agent diamide, the index was 47.0 +/- 2.6 (pH 7.4). In the CL-Cyt c-PLA2 system, lipid peroxidation seemed to be due to three mechanisms: (1) oxidation of the LA (nonreleased form) constituent of CL by Cyt c (oxidation of CL by Cyt c); (2) oxidation of free LA, released from CL, involving the oxidation of CL by Cyt c (free LA oxidation by the CL-Cyt c complex); and (3) oxidation of free LA, released from CL, by Cyt c and calcium ions (LA-Cyt c-Ca system). The lipid peroxidation of the CL-Cyt c-PLA2 system was also enhanced by the addition of diamide and by an acidic pH of 6.7. The fact that the SH-oxidizing agent enhanced the lipid peroxidation in the CL-Cyt c-PLA2 system suggested that SH groups in the hemoprotein played an inhibitory role in lipid peroxidation in the system.

We report two infant deaths attributable to intussusception, but without clear evidence of peritonitis. In the first instance, a 3-year-old girl had presented with abdominal pain, vomiting, and melena before her demise. Aspirated vomitus was subsequently ascertained as the immediate cause of death, due to intussusception-induced ileus. The other infant, a 2-month-old male, showed autopsy evidence of intussusception at two sites, with likely aspiration of gastric mucus. Since the circumstances surrounding his death were vague, timing of the intussusception was difficult to pinpoint. Thus, an inconsequential, agonal event could not be discounted. Taken together, however, death from intussusception, without peritonitis, is the most viable postmortem interpretation for both patients. The causes of death in such cases are established by comprehensive delineation of preceding clinical events, plus autopsy documentation of coexistent intussusception and vomitus aspiration.

Pink teeth is thought to result from the seepage of hemoglobin caused by dental pulp decomposition. We investigated whether racemization can be applied for age estimation in cases of pink teeth where the whole tooth is used. The pink teeth used were three cases and the normal teeth for control were five mandibular canines of known age. Age of the pink teeth was calculated on the basis of regression formula obtained from the five control teeth. Only a slight error was noted between the actual and estimated ages of the pink teeth (R(2) = 0.980, r = 0.990): Cases 1-3 actually aged 23, 53, and 59 years were estimated to be 26, 52, and 60 years. Based on our results of testing pink teeth of known age, we suggest that racemization techniques allow for the age estimation of pink teeth using the same methods for normally colored teeth.

IgG4-related disease (IgG4-RD) is a systemic inflammatory disease characterized by marked infiltration of IgG4-positive (+) plasma cells into affected organs, but the concept of this disease has only recently been established. Coronary vasculitis is a rare disease that can cause sudden death, and it has recently been reported that IgG4-RD may be associated with vasculitis, including periarteritis and coronary disease. In this paper we report an autopsy case of sudden death of a man in his thirties, in which coronary periarteritis with features of IgG4-related periarteritis was detected. IgG4-RD was suspected from the presence of the following histopathological features: (1) markedly thickened adventitia and marked infiltration of the adventitia and periarterial fat by lymphocytes and plasma cells; and (2) infiltration of IgG4-positive plasma cells (ratio of IgG4+ cells to IgG4+ cells of >40 %, 50 IgG4+ plasma cells per high-power field) on immunostaining. The etiology and pathophysiology of IgG4-RD and IgG4-related periarteritis are still unclear, and further investigation of these conditions and their association with coronary lesions is needed. Careful consideration should be given to the possible presence of IgG4-RD when forensic pathologists encounter cases of sudden death accompanied by coronary periarteritis.

Superfusion with a cyclopropane fatty acid, cis-9, 10-methylenehexadecanoic acid (10-300 microM), reduced the contractility of papillary muscle isolated from guinea pigs in a dose-dependent manner. cis-9,10-Methylenehexadecanoic acid also inhibited the Mg(2+)-ATPase activity of guinea pig papillary myocardium by about 40% at 400 microM. Since cis-9, 10-methylenehexadecanoic acid 4 microM inhibited the K(+)-EDTA-ATPase activity inherent in myosin's catalytic activity by about 25%, the fatty acid was thought to interact with the catalytic center of the myosin molecule.

Cytochrome c oxidase (COX), a mitochondrial enzyme, is inactivated by cyanide or carbon monoxide (CO) intoxication. To test whether cytochrome c has potential as an indicator of these toxins in cadavers, we measured COX activity in the main organs of the rat, and in the human heart, at various times after death. Each tissue sample or organ was homogenized and the COX activity in the mitochondrial fraction was measured using ferrous cytochrome c as the substrate. COX activity was significantly higher in rat brain, heart and kidney than in lung and liver from 0 to 4 days after death. The loss of COX activity was significantly slower in the brain and heart than in the lung, liver and kidney. Most importantly, COX activity correlated with the time-since-death for each of the rat organs we tested (r2=0.70-0.95), but for the human heart (r2=0.47). It may be possible that COX activity is likely to be a useful indicator of the time-since-death, and is worth pursuing as an indicator of the tissue cyanide and CO content.

The DNA of aortic tissues collected at the autopsies of unidentified 47 cadavers was examined using a multiplex short tandem repeat (STR) typing kit. The causes of death included drowning, burning and brain injury among others. Tissues samples were stored in ethanol before DNA extraction. DNA was extracted from about 25mg of dried tissues using a Q1Amp DNA Mini kit (QIAGEN). STR typing was performed using an AmpFlSTR Identifiler PCR Amplification kit (Applied Biosystems) and GeneMapper ID software v. 3.2 (Applied Biosystems). The amount of recovered DNA ranged from 0.006 to 3.44 microg/mg. Tissue samples were collected at estimated times between 1 day and 2 years after death. We were able to type 46/47 tissue samples (98%) and all 15 STR alleles and the amelogenin gene were detected in 38 cases (81%). Successful typing was completed for most tissue samples taken less than 1 month and up until 3 months after death. As the days after death increased, the numbers of alleles with longer DNA fragment sizes decreased. These results suggest that the DNA from aortic tissues can be accurately typed for multiplex STR and amelogenin until about 1 month after death. We found that aortic tissues are one of the most useful samples for forensic personal identification of unidentified bodies.

A corpse completely converted into adipocere and showing two adjacent bone defects—a typical gunshot entrance wound and a keyhole lesion—is reported. Postmortem changes, a comminuted fracture of the cranial base, and destruction of the bullets made it impossible to determine the direction of fire through the keyhole lesion. Gunshot wounds may show various atypical forms, including keyhole lesions, and especially in old corpses, the distinction between an entrance wound and an exit wound can be very difficult, even if a careful complete autopsy is performed.

We examined the postmortem changes in the levels of ATP, glycogen and lactic acid in two masticatory muscles and three leg muscles of rats. The proportion of fibre types of the muscles was determined with NIH image software. The ATP levels in the white muscles did not decrease up to 1 h after death, and the ATP levels 1 and 2 h after death in the white muscles were higher than those in the red muscles with a single exception. The glycogen level at death and 1 h after death and the lactic acid level 1 h after death in masticatory muscles were lower than in the leg muscles. It is possible that the differences in the proportion of muscle fibre types and in glycogen level in muscles influences the postmortem change in ATP and lactic acid, which would accelerate or retard rigor mortis of the muscles.

We report that there is a time-related change in the phospholipase C (PLC) activities of rat brain cytosol and membrane fractions after iv injection of a soman-like or a sarin-like organophosphorous agent (bis(isopropyl methyl)phosphonate [BIMP] and bis(pinacolyl methyl)phosphonate [BPMP]). PLCgamma was activated in the brain cytosol fraction from BPMP-injected rats. The phosphorylating activity of rat brain membrane fractions were enhanced by BPMP treatment. The brain membrane fractions from BPMP-treated rats phosphorylated several proteins, including supposedly PLCgamma in the brain cytosol fraction from control rats in vitro. These results suggest that soman and sarin may stimulate a membrane tyrosine kinase, including growth factor receptors, directly or indirectly.

We report a case of subarachnoid hemorrhage at the skull base with possible rupture of a vertebral artery, and we present a new method to investigate rupture of this artery. In this method, the brain stem and cerebrum are divided with ligation of the internal carotid artery and basilar artery, and milk is injected from the original part of the vessel to find the rupture. We show that this method is particularly useful for cases of subarachnoid hemorrhage at the skull base in which there may be rupture of a vertebral artery. Language: en

Bovine and guinea pig heart homogenates, porcine leukocyte homogenate, and human hemolysate were found to vigorously oxidize linoleic acid, with lipoxygenase-like activity, to its hydroperoxy, epoxy, hydroxy-epoxy, and keto compounds in the presence of calcium chloride. In the absence of calcium, the reaction was significantly reduced. Attempts to characterize this quasi-lipoxygenase activity revealed that calcium potentiated the quasi-lipoxygenase activities of hemoproteins (hemoglobin, myoglobin, myeloperoxidase, catalase, cytochrome c) and hemin at the physiological pH of 7.5. Lipid peroxidation by hemoproteins was inhibited by albumin and erythrocyte membranes in blood, as well as by a low concentration of calcium in cells. However, it seems possible that in extracellular fluid, which contains a high concentration of calcium and a low concentration of albumin, hemoprotein released from damaged cells could oxidize unsaturated fatty acids derived by phospholipase-A2 from phospholipids of damaged cellular membranes. In a model of quasi-lipoxygenase activation under such conditions, lipids of erythrocyte membranes were oxidized by hemoglobin in the presence of phospholipase-A2 and calcium. The effect of nitrogen oxide, paraquat, and bleomycin on oxidation by hemoproteins and hemin was also discussed.

To observe lipid peroxidation of additive-free submitochondrial particles, we incubated submitochondrial particles in the absence of exogenous irons and t -butyl hydroperoxide. After the incubation, the phospholipids were hydrolyzed by phopholipase A 2 , and the fatty acid constituents were analyzed by high-performance liquid chromatography, gas chromatography-mass spectrometry, and liquid chromatography-mass spectrometry. Contrary to a commonly accepted theory, lipid peroxidation in the submitochondrial particles did not need the addition of NADH. In the phospholipid constituent fatty acids of the oxidized submitochondrial particles, derivatives of hydroperoxides of linoleic acid such as keto, hydroxy, trihydroxy, and hydroxyepoxy compounds were generated. Lipid peroxidation in the submitochondrial particles was not inhibited by the addition of catalase, superoxide dismutase, hydroxyl radical scavengers, or ethylenediaminetetraacetic acid, but was inhibited by the addition of KCN, antimycin-A, NADH, ubiquinol, deferoxamine mesylate, ascorbic acid, and α-tocopherol. The cardiolipin–cytochrome c lipid peroxidation system could mimic the lipid peroxidation of the submitochondrial particles, in terms of linoleic acid products and the inhibitory patterns of radical scavengers and electron transfer chain inhibitors. Thus, lipid peroxidation in the submitochondrial particles seems to be due to phospholipid–hemoprotein lipid peroxidation systems such as the cardiolipin–cytochrome c system.

One of the hydrolysis products of sarin (isopropyl methylphosphonofluoridate) was detected in formalin-fixed brain tissues of victims poisoned in the Tokyo subway terrorist attack. Part of this procedure, used for the detection of sarin hydrolysis products in erythrocytes of sarin victims, has been described previously. The test materials were four individual cerebellums, which had been stored in formalin fixative for about 2 years. Sarin-bound acetylcholinesterase (AChE) was solubilized from these cerebellums, purified by immunoaffinity chromatography, and digested with trypsin. Then the sarin hydrolysis products bound to AChE were released by alkaline phosphatase digestion, subjected to trimethylsilyl derivatization (TMS), and detected by gas chromatography-mass spectrometry. Peaks at m/z 225 and m/z 240, which are indicative of TMS-methylphosphonic acid, were observed within the retention time range of authentic methylphosphonic acid. However, no isopropyl methylphosphonic acid was detected in the formalin-fixed cerebellums of these 4 sarin victims, probably because the isopropoxy group of isopropyl methylphosphonic acid underwent chemical hydrolysis during storage. This procedure will be useful for the forensic diagnosis of poisoning by protein-bound, highly toxic agents, such as sarin, which are easily hydrolysed. This appears to be the first time that intoxication by a nerve agent has been demonstrated by analyzing formalin-fixed brains obtained at autopsy.

A sarin-like organophosphorus agent, [bis(isopropyl methyl)phosphonate; BIMP], was synthesized. This agent has the same phosphonate group as sarin and also has the same anti-acetylcholinesterase activity potency as sarin. The ID50 and LD50 values of BIMP in mice after intravenous injection were 3.9 nM and 0.8 mg/kg, respectively. The AChE activities of their red blood cells and brains were dose-dependently reduced by intravenous BIMP. After preparation of experimental BIMP-exposed human red blood cells, BIMP-bound acetylcholinesterase (AChE) was solubilized from erythrocyte membranes, purified by immunoaffinity chromatography, digested with trypsin, and the sarin hydrolysis products bound to AChE were released by alkaline phosphatase digestion. The digested sarin hydrolysis products were subjected to trimethylsilyl (TMS) derivatization and detected by gas chromatography-mass spectrometry. Isopropyl methylphosphonic- and methylphosphonic acids, which are the sarin hydrolysis products, were detected in experimental BIMP-exposed human red blood cells. This new method, which enables sarin's hydrolysis products to be detected in BIMP-exposed erythrocytes, is a useful tool for studying sarin-poisoning victims.

Studies of four male victims who were killed in an accidental tunnel gas explosion on urban reclaimed land are described. The studies were judicial autopsy examinations to determine the precise causes of death. Two men died of carbon monoxide intoxication, one died of massive brain damage, and the fourth died of drowning. The concentrations of methane in several organs were much lower than the lethal level, whereas those in adipose tissue were relatively high. These findings indicated that a low concentration of methane was almost always present in the atmosphere at the construction site. Recently, coal mine accidents have been decreasing in Japan. However, there is still a possibility of underground explosions or gas leaks in confined spaces other than coal mines. To determine the precise cause of death in such cases, careful autopsies and other examinations should be performed using methods similar to those used in coal mine accidents.

We used a mobile computed tomography (CT) unit for postmortem examinations of deceased subjects to see how many mistakes on cause-of-death diagnoses were made in Japan. In 5 of 20 cases, the cause of death determined by CT was different from the diagnosis made by superficial postmortem examination. In one case, the superficial examination suggested no trauma, whereas a subdural hematoma was found on cranial CT images. We concluded that postmortem examinations in Japan were not effective when screening for crimes or accidents. Using a mobile CT scanner in postmortem examination may be a viable method of screening for causes of deaths, although it cannot be used as a substitute for autopsy.

A man in his twenties was diagnosed with schizophrenia in his late teens. The night before his death, his family reported he drank a large amount of water, vomited, collapsed, and snored loudly while sleeping, but they did not view the event seriously as he did it routinely. The following morning, he was found dead. Autopsy revealed hyponatremia by water intoxication as the cause of death. Water intoxication has various causes. In this case, 610 ng/mL olanzapine was detected in serum samples. Although this concentration is not as high as the fatal concentrations reported in past studies, it might have caused some adverse effects. Furthermore, the observation that excessive drinking behavior started after the dose of olanzapine was increased suggests a possibility that olanzapine aggravated water intoxication.

We previously reported that superimposition of 3-dimensional (3D) images reconstructed from computed tomographic images of skeletonized skulls on photographs of the actual skulls afforded a match of skull contours, thereby demonstrating that superimposition of 3D-reconstructed images provides results identical to those obtained with actual skulls. The current superimposition procedure requires a skeletonized skull with mouth closed and thus is not applicable to personal identification using a skull with residual soft tissue or the mouth fixed open, such as those found in mummified or burned bodies. In this study, we scanned using computed tomography the skulls of mummified and immersed body with mandibles fixed open by residual soft tissue, created 3D-reconstructed skull images, which were digitally processed by computer software to close the mandible, and superimposed the images on antemortem facial photographs. The results demonstrated morphological consistency between the 3D-reconstructed skull images and facial photographs, indicating the applicability of the method to personal identification.

The novel cytotoxic factor Subtilase cytotoxin (SubAB) is produced mainly by non-O157 Shiga-toxigenic Escherichia coli (STEC). SubAB cleaves the molecular chaperone BiP/GRP78 in the endoplasmic reticulum (ER), leading to activation of RNA-dependent protein kinase (PKR)-like ER kinase (PERK), followed by caspase-dependent cell death. However, the SubAB uptake mechanism in HeLa cells is unknown. In this study, a variety of inhibitors and siRNAs were employed to characterize the SubAB uptake process. SubAB-induced BiP cleavage was inhibited by high concentrations of Dynasore, and methyl-β-cyclodextrin (mβCD) and Filipin III, but not suppressed in clathrin-, dynamin I/II-, caveolin1- and caveolin2-knockdown cells. We observed that SubAB treatment led to dramatic actin rearrangements, e.g., formation of plasma membrane blebs, with a significant increase in fluid uptake. Confocal microscopy analysis showed that SubAB uptake required actin cytoskeleton remodeling and lipid raft cholesterol. Furthermore, internalized SubAB in cells was found in the detergent-resistant domain (DRM) structure. Interestingly, IPA-3, an inhibitor of serine/threonine kinase p21-activated kinase (PAK1), an important protein of macropinocytosis, directly inhibited SubAB-mediated BiP cleavage and SubAB internalization. Thus, our findings suggest that SubAB uses lipid raft- and actin-dependent, but not clathrin-, caveolin- and dynamin-dependent pathways as its major endocytic translocation route.

Historically, coronary angiography of the isolated heart has played an important role in the detection of stenotic or occlusive lesions that are difficult to identify by autopsy alone. Meanwhile, although the application of multidetector computed tomography (MDCT) to forensic fields has accelerated recently, isolated single organ angiography with MDCT is rarely performed. In this article, we present an evaluation of postmortem selective coronary CT angiography of the isolated heart with MDCT and discuss its utility for autopsy. First, in a preliminary experiment using pig coronary artery, we examined the behavior of water soluble contrast material on postmortem computed tomography angiography (PMCTA) and found that better angiographic images were acquired when the viscosity of the contrast material was increased and CT was performed under conditions of sustained perfusion. Based on these results, we devised a selective coronary angiography procedure using a pressurized bag for drip infusion that can be performed easily, quickly, and at low cost. The angiographic images obtained provided useful supportive evidence of autopsy findings suggestive of ischemic heart disease. With active discussions underway in forensic fields on the proper use of postmortem computed tomography, PMCTA has also naturally attracted attention as it compensates for some of the shortcomings of CT alone. Although PMCTA typically involves whole-body angiography, if we view PMCTA as one of the many useful and supplementary tools available for autopsy, then isolated heart angiography continues to have utility in autopsy today.