Estimating age at death by Hausdorff distance analyses of the fourth lumbar vertebral bodies using 3D postmortem CT images.

The existing methods for determining adult age from human skeletons are mostly qualitative. However, a shift in quantifying age-related skeletal morphology on a quantitative scale is emerging. This study describes an intuitive variable extraction technique and quantifies skeletal morphology in continuous data to understand their aging pattern. A total of 200 postmortem CT images from the deceased aged 25-99 years (130 males, 70 females) who underwent forensic death investigations were used in the study. The 3D volume of the fourth lumbar vertebral body was segmented, smoothed, and post-processed using the open-source software ITK-SNAP and MeshLab, respectively. To measure the extent of 3D shape deformity due to aging, the Hausdorff distance (HD) analysis was performed. In our context, the maximum Hausdorff distance (maxHD) was chosen as a metric, which was subsequently studied for its correlation with age at death. A strong statistically significant positive correlation (P < 0.001) between maxHD and age at death was observed in both sexes (Spearman's rho = 0.742, male; Spearman's rho = 0.729, female). In simple linear regression analyses, the regression equations obtained yielded the standard error of estimates of 12.5 years and 13.1 years for males and females, respectively. Our study demonstrated that age-related vertebral morphology could be described using the HD method. Moreover, it encourages further studies with larger sample sizes and on other population backgrounds to validate the methodology.

Rescue from Stx2-Producing E. coli-Associated Encephalopathy by Intravenous Injection of Muse Cells in NOD-SCID Mice.

Shiga toxin-producing Escherichia coli (STEC) causes hemorrhagic colitis, hemolytic uremic syndrome, and acute encephalopathies that may lead to sudden death or severe neurologic sequelae. Current treatments, including immunoglobulin G (IgG) immunoadsorption, plasma exchange, steroid pulse therapy, and the monoclonal antibody eculizumab, have limited effects against the severe neurologic sequelae. Multilineage-differentiating stress-enduring (Muse) cells are endogenous reparative non-tumorigenic stem cells that naturally reside in the body and are currently under clinical trials for regenerative medicine. When administered intravenously, Musecells accumulate to the damaged tissue, where they exert anti-inflammatory, anti-apoptotic, anti-fibrotic, and immunomodulatory effects, and replace damaged cells by differentiating into tissue-constituent cells. Here, severely immunocompromised non-obese diabetic/severe combined immunodeficiency (NOD-SCID) mice orally inoculated with 9 × 109 colony-forming units of STEC O111 and treated 48 h later with intravenous injection of 5 × 104 Muse cells exhibited 100% survival and no severe after-effects of infection. Suppression of granulocyte-colony-stimulating factor (G-CSF) by RNAi abolished the beneficial effects of Muse cells, leading to a 40% death and significant body weight loss, suggesting the involvement of G-CSF in the beneficial effects of Muse cells in STEC-infected mice. Thus, intravenous administration of Muse cells could be a candidate therapeutic approach for preventing fatal encephalopathy after STEC infection.

Autopsy case of sudden maternal death from thrombotic thrombocytopenic purpura.

A 31-year-old pregnant woman was transferred to the emergency room at 27 weeks of gestation. She had one-day history of fever and upper abdominal pain. Soon after admission, she underwent cardiopulmonary arrest. Autopsy was performed and multiple microthrombi were seen within the small-caliber vessels of many organs, but not in the lungs. Immunohistochemical staining revealed that the thrombi were rich in von Willebrand factor. We also obtained results which showed severely deficient plasma a disintegrin-like and metalloprotease with thrombospondin motifs (ADAMTS) 13 activity and positive ADAMTS13 inhibitor, confirming a diagnosis of thrombotic thrombocytopenic purpura. As far as we know, in Japan, this is the first autopsy report of sudden maternal death from thrombotic thrombocytopenic purpura. We expect that the routine laboratory application of ADAMTS13 assays for unknown thrombocytopenic patients during pregnancy may help in differential diagnosis at an earlier stage of the disease and facilitate tailor-made therapeutic intervention.

Vascular anatomy and their variations in Situs inversus totalis using postmortem computed tomographic angiography.

Studies describing the vascular systems and their variations in Situs inversus totalis (SIT) from a whole-body computed tomographic (CT) angiography perspective are lacking. We report a case of SIT in which postmortem CT angiography (PMCTA) was performed as a part of the forensic death investigation and incidentally detected several vascular variations in it. The PMCTA procedure was performed using the multiphase PMCTA protocol. Almost all major vessels were visualized, indeed in a completely reversed pattern. Contrast mixture flow interruptions were noted in the right coronary arterial branches suggesting possible blockage, upon which autopsy revealed >90% vessel occlusions at several locations. As such the cause of death was due to ischemic heart disease. Anomalous origins of the right internal mammary artery; abnormal left thyrocervical trunk and variations in the drainage of testicular veins were noted. Our findings might be helpful to clinicians and add to the body of literature on SIT.

Development of New Formulas for Sex and Age Estimation by Assessing Maxillary Sinus Morphology on CBCT.

Background: The morphology of the maxillary sinus varies between individuals which could be used in the forensic personal identification process. Methods: In the current study, the CBCT images of the maxillary sinus in 453 patients (217 males, 236 females) aged 14 to 95 years were analyzed. In particular, each left, and right maxillary sinus of the subjects was measured for its maximum height, width, and breadth in 2-D, and volume in 3-D perspectives, and their usefulness for age and sex estimation was examined. Regarding age estimation, because the size of the maxillary sinus increases up to 20s and then decreases over time, two separate age estimation formulas were created, one for subjects in their 14-21 years and the other for those over 22 years old. For each age group, multiple regression formulas were generated using the diameters and volume as explanatory variables and the chronological age as a response variable. This study used 150 cases not included in the study as a validation set for age estimation. Results: Generally, all the diameters and volumes in both sinuses tended to increase till the mid-20s, and then gradually decreased over time. The derived formulas were tested for their accuracy on additional 150 subjects. Plausibly, the model could estimate the age between 14-21 years old with an average accuracy of ± 1.8 years for men and ± 3.2 years for women. Whereas for those over 22 years old, it was possible to estimate the age with an accuracy of ± 11.8 years for males and ± 10.3 years for females, respectively. A comparison of estimated age and chronological age did not show a statistically significant difference(P > 0.05). It was found that the left maxillary sinus had more age groups showing the most significant difference than other measurements between sexes(P < 0.05). The maxillary sinus height may be significantly affected by gender differences. Conclusion: Overall, this study showed the effectiveness of age and sex estimation using the maxillary sinus morphometric analyses.

A novel method to estimate adult age from the lumbar vertebral body using 3D PMCT images in Japanese.

This study evaluated the age-related changes in the vertebral body using 3D Postmortem CT (PMCT) images and proposed an alternative age estimation formula. The PMCT images of 200 deceased individuals aged 25 to 99 years (126 males, 74 females) were retrospectively reviewed and included in the study. Using the open-source software ITK-SNAP and MeshLab, a 3D surface mesh of the fourth lumbar vertebral body (L4) and its convex hull models were created from the PMCT data. Using their inbuilt tools, volumes (in mm3) of the L4 surface mesh and convex hull models were subsequently computed. We derived VD, defined as the difference in volumes between the convex hull and L4 surface mesh normalized by L4 mesh volume, and VR, defined as the ratio of L4 mesh volume to convex hull volume based on individual L4. Correlation and regression analyses were performed between VD, VR, and chronological age. A statistically significant positive correlation (P < 0.001) between chronological age and VD, (rs = 0.764, males; rs = 0.725, females), and a significant negative correlation between chronological age and VR (rs = -0.764, males; rs = -0.725, females) was obtained in both sexes. The lowest standard error of the estimate was demonstrated by the VR at 11.9 years and 12.5 years for males and females, respectively. As such, their regression models to estimate adult age were Age = 248.9-2.5VR years, males; Age = 258.1-2.5VR years, females. These regression equations may be useful for estimating age in Japanese adults in forensic settings.

Forensic Medicine in South Asia: Comparison to the Developed Countries.

This study attempted to discuss the historical context and current practice of forensic medicine in South Asia. Comparisons within and between countries in South Asia, and between South Asia and the developed countries (represented by Japan and the USA) have been made to provide an insight into their distinct practice of forensic medicine. Though the formal establishment of forensic medicine in South Asia commenced at a comparable period to the developed countries, their pace of development has been considerably slow. Moreover, their ways of practice as well have evolved differently. In effect, South Asian countries follow an 'integrated service' system, whilst Japan and the USA practice 'divided service' systems to provide forensic medical services. Similarly, regarding the death investigations, most South Asian countries follow a Police-led death investigation system, whereas Japan and the USA follow a hybrid model and the Medical examiner's system of death investigation, respectively. Indeed, forensic medicine in South Asia is undeniably underdeveloped. In this paper, by highlighting the issues and challenges confronted in South Asia, key actions for prompt redressal are discussed to improve the standard of forensic medical services in South Asia.

Estimation of Postcardiac Arrest Interval Based on Atrial Cavity Density in Postmortem Computed Tomography.

BACKGROUND: Intracardiac hypostasis is frequently observed in postmortem computed tomography (PMCT) of acute deaths, and it becomes clearer as the postmortem interval increases. To determine the postcardiac arrest interval (PCAI), we evaluated densities of the right and left atria [anterior part of the right atrium (AR) and posterior part of the left atrium (PL)] using postmortem computed tomographic images. METHODS: A total of 184 patients were included in the study. Patients with return of spontaneous circulation and those with final alive confirmation time over 1 hour were excluded. We evaluated the density of AR and PL at the level of the right inferior pulmonary vein entry to the left atrium. We defined the interval between the estimated cardiac arrest time and the postmortem CT time as the PCAI. RESULTS: There was a negative correlation between AR and PCAI in 59 patients who died owing to cardiovascular disease. The regression equation (PCAI = -1.725 × AR + 132.95) was obtained based on this result. There was no correlation between PL and PCAI. CONCLUSION: The result suggests that the density of the anterior part of the right atrium decreases as postcardiac arrest interval increases in the case of cardiovascular disease. The regression equation may be used as an additional method to estimate postcardiac arrest interval.

Expression of toll-like receptors in the pancreas of recent-onset fulminant type 1 diabetes.

Fulminant type 1 diabetes, established in 2000, is defined as a novel subtype of diabetes mellitus that results from remarkably acute and almost complete destruction of pancreatic beta cells at the disease onset. In this study, we aimed to clarify the pathogenesis of fulminant type 1 diabetes with special reference to insulitis and viral infection. We examined pancreatic autopsy samples from three patients who had died soon after the onset of disease and analyzed these by immunohistochemistry and in situ-hybridization. The results were that both beta and alpha cell areas were significantly decreased in comparison with those of normal controls. Mean beta cell area of the patients just after the onset was only 0.00256 % while that of normal control was 1.745 %. Macrophages and T cells-but no natural killer cells-had infiltrated the islets and the exocrine pancreas. Although both of them had massively infiltrated, macrophages dominated islet infiltration and were detected in 92.6 % of the patients' islets. Toll-like receptor (TLR) 3, a sensor of viral components, was detected in 84.7+/- 7.0 % of T cells and 62.7+/- 32.3 % of macrophages (mean+/- SD) in all three patients. TLR7 and TLR9 were also detected in the pancreas of all three patients. Enterovirus RNA was detected in beta-cell positive islets in one of the three patients by in situ-hybridization. In conclusion, our results suggest that macrophage-dominated insulitis rather than T cell autoimmunity contributes to beta cell destruction in fulminant type 1 diabetes.

Minisatellite MS32 alleles show population specificity among Thai, Chinese, and Japanese.

Lineages of structurally related alleles at minisatellite MS32 in human populations show considerable differentiation at the continental level. However, the regional specificity of these lineages remains unknown. We now describe the comparison of allele structures in Thai, Han Chinese, and Japanese populations with lineages previously established for North Europeans and Africans. The great majority of alignable Asian alleles showed their closest structural relative in Asia, with few instances of preferential alignment of Asian with European alleles and only one isolated incident showing a best match with an African allele. Further, there was a strong tendency, most marked for Japanese, for Asian alleles to align preferentially with other alleles from the same population, indicating strong regional specificity of allele lineages. This rapidly evolving minisatellite can therefore serve as a lineage marker for exploring recent events in human population history and dissecting population structure at the fine-scale level, as well as being an extremely informative DNA marker for personal identification.

Japaneseplex: A forensic SNP assay for identification of Japanese people using Japanese-specific alleles.

It is sometimes necessary to determine whether a forensic biological sample came from a Japanese person. In this study, we developed a 60-locus SNP assay designed for the differentiation of Japanese people from other East Asians using entirely and nearly Japanese-specific alleles. This multiplex assay consisted of 6 independent PCR reactions followed by single nucleotide extension. The average number and standard deviation of Japanese-specific alleles possessed by an individual were 0.81 ±â€¯0.93 in 108 Koreans from Seoul, 8.87 ±â€¯2.89 in 103 Japanese from Tottori, 17.20 ±â€¯3.80 in 88 Japanese from Okinawa, and 0 in 220 Han Chinese from Wuxi and Changsha. The Koreans had 0-4 Japanese-specific alleles per individual, whereas the Japanese had 4-26 Japanese-specific alleles. Almost all Japanese were distinguished from the Koreans and other people by the factorial correspondence and principal component analyses. The Snipper program was also useful to estimate the degree of Japaneseness. The method described here was successfully applied to the differentiation of Japanese from non-Japanese people in forensic cases. This Japanese-specific SNP assay was named Japaneseplex.

Unexpected sudden death resulting from anomalous origin of the right coronary artery from the left sinus of Valsalva: a case report involving identical twins.

An anomalous origin of the coronary arteries had been implicated as a cause of sudden cardiac death in a case involving a 16-year-old student who suddenly collapsed while running track at school. An autopsy revealed that the right coronary artery arose from the aorta in the left sinus of Valsalva. In order to determine whether the same anomaly was inherent in his brother--an identical twin--a complete cardiac medical examination was conducted. Multi-detector-row computed tomography (MDCT) coronary angiography showed no anomaly at the sinus of Valsalva, which indicates that the anomaly, in this case, was not hereditary. This case exemplifies instances where forensic medicine may intervene to prevent sudden deaths in surviving family members.

Genotyping of the c.1423C>T (p.P475S) polymorphism in the ADAMTS13 gene by APLP and HRM assays: Northeastern Asian origin of the mutant.

ADAMTS13 is a von Willebrand factor-cleaving protease. The mutant types of p.P475S (c.1423C>T) polymorphism in ADAMTS13 have a reduced activity in comparison with the wild type. In the present study, we investigated the frequency of the C-to-T substitution in 2584 genomic DNA samples from 25 Asian, European, and African populations using APLP (amplified product length polymorphism) and/or HRM (high-resolution melting) assays. Allele T (ADAMTS13(∗)T) was detected only in Asian populations and its frequency was observed to decrease gradually from north to south in 24 East Asian populations. Almost all ADAMTS13(∗)T were associated with ABO(∗)O. These results suggested that ADAMTS13(∗)T had occurred on a chromosome with ABO(∗)O in a northern part of East Asia. This SNP is useful as an ancestry-informative marker, and the present genotyping techniques are applicable to the investigation of an association between this SNP and aortic dissection (Kobayashi et al., 2012).

Maternal methamphetamine administration during pregnancy influences on fetal rat heart development. [corrected].

Methamphetamine (MAP) is one of the most abused drugs in Japan. The rate of MAP abuse by young women has recently reached more than 50 percent in adolescents. A major health concern is that these women will continue to use MAP during pregnancy. The purpose of this study was to investigate whether MAP administered to the mother during pregnancy would change the expression of alpha- and beta- myosin heavy chain (MHC) mRNA in rat neonatal hearts, as detected by quantitative RT-PCR. In addition, morphological changes in the rat neonatal ventricles were examined. Pregnant rats were injected intraperitoneally with MAP (1 mg/kg/day) starting at day 0 of gestation and ending at day 21. There was a significant increase in alpha-MHC mRNA expression in the neonatal ventricular muscle in the experimental group compared with the control at postnatal day (P) 0 and 5. alpha-MHC mRNA expression in both groups was similar after P9. beta-MHC mRNA expression was similar in both groups at P0. Postnatal beta-MHC mRNA expression decreased rapidly, but significant alteration was not detected. Neonatal rats at P0 exhibited some cardiac changes, including hypertrophy, degeneration, and disarrangement of myofibers, but these lesions disappeared by P14. We conclude that chronic maternal administration of MAP changes the alpha- and beta-MHC mRNA expression pattern in fetal and neonatal hearts, correlating with abnormal development, plasma level of hormones, and myocardial damage. At the same time, it is indicated that neonatal cardiomyocytes have reversibility.

Obesity and sudden unexpected deaths in Osaka, Japan.

Obesity and cardiomegaly/hypertension may be strongly associated with sudden unexpected deaths (SUD) due to circulatory diseases. Six hundred and forty-nine SUD involving 402 postmortems, aged between 10 and 59 years in Osaka in 1997 were analyzed using the calculated body mass index (BMI) and the calculated degree of cardiac hypertrophy (DCH) by Hitosugi (Legal Med 1999;1:80). The percentage of individuals who died due to circulatory diseases was 54% in men and 64% in women, and at ages 50-59 years, 60% in men and 75% in women. It was 80% with DCH>/=20%, 84% for individuals with hypertension as a past illness and 80% with BMI>/=24. Thirty-four percent of all SUD have cardiomegaly more than 20%, 41% have BMI more than 24, and 17% have at least hypertension as a past illness.

Simultaneous determination of flunitrazepam and 7-aminoflunitrazepam in human serum by ion trap gas chromatography-tandem mass spectrometry.

A method for the determination of flunitrazepam (FNZ) and 7-aminoflunitrazepam (7-AFNZ) in human serum was developed with ion trap gas chromatography (GC)-tandem mass spectrometry. The 7-AFNZ was derivatizated with 50 microl trifluoroacetic anhydride (TFAA), 60 degrees C-20 min. EI mass spectra and tandem mass spectra of FNZ and 7-AFNZ-TFA were m/z 238, 239, 266, 286, 294, 312, 313(M(+)), m/z 350, 351, 360, 378, 379(M(+)), m/z 238, 239, 240 (precursor ion m/z 286, collision energy 1.5 V), and m/z 239, 254, 264, 336 (precursor ion m/z 351, collision energy 1.8 V), respectively. The detection limits of full scan EI mass spectrometry and tandem mass spectrometry for FNZ and 7-AFNZ in human serum were ca. 200 ng/ml, 60 ng/ml, 15 ng/ml and 1 ng/ml, respectively.

A case of an infant suffered from a fatal scald injury - validity and limitations of postmortem CT imaging.

A 9months-old infant has died of scald burn by the hot water from the fallen electric pot at home. Postmortem computed tomography (CT) scanning prior to autopsy contributed to exclude the possibilities of old and new bone fractures or hidden massive hemorrhages. This procedure helped the pathologists to avoid dissecting all the extremities, which is time-consuming, laborious and often hurtful to the innocent parents of the deceased. On the other hand it did not successfully show the distribution of the scald burn injury on the skin, which reminded us of the importance of external examination even when the whole-body CT is performed. The manner of death was considered to be accident since the cause of death was burn shock and there was no significant evidence suggesting child abuse. Scene investigation and reproductive experiment have revealed that there was a structural problem in the lid of the pot, which comes off very easily in a certain condition. In order to prevent a similar accident, report of the case to the manufacturer is essential for the improvement of the apparatus. This case, however, has been remained to be unreported because of the confidentiality of the investigative information in judicial autopsy. Finally, we recommend a partial revision of the reporting system in Japan to prevent the recurrence of such tragic accidents.

Latent adrenal Ewing sarcoma family of tumors: A case report.

Ewing sarcoma family of tumors (ESFT) is derived from the neural crest, which originates from basal embryo cells in the primitive neural tube. ESFT often arises at the bone, chest wall, and soft tissues of the thoracic region. However, ESFT that arises from the adrenal gland is much rarer and it is usually revealed by clinical symptoms. We report an autopsy case of suicidal hanging, in which adrenal ESFT was incidentally revealed. To our knowledge, this is the first case of latent ESFT arising from the adrenal gland. Autopsy can sometimes reveal latent disease. Some of these latent diseases are very rare and we would not be able to detect them without a complete autopsy. As forensic pathologists, we should attempt to perform a complete autopsy and report new discoveries for the development of medicine.

Postmortem computed tomography findings of upper airway obstruction by food.

This study is a retrospective analysis of 14 cases with food bolus upper airway obstruction as the defined cause of death where both postmortem computed tomography and autopsy were performed. Three groups were defined by the images i.e., Type 1: foreign body situated between the oral cavity and oropharynx, while the epiglottis sits in normal position, Type 2: foreign body situated in the oropharynx just above the epiglottis pushing it posteriorly and obstructing the airway, and Type 3: foreign body obstructing the laryngeal inlet while pushing the epiglottis anteriorly. At the time of autopsy, foreign bodies were detected by pathologists, occasionally in a different position, presumably being dislodged in the act of organ removal especially for the "Type 1" pattern. CT imaging provides accurate interrogation of upper airway bolus obstruction prior to autopsy.