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.

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.

Y chromosome analysis for common surnames in the Japanese male population.

For many years of Japan's long history, Japanese surnames have been handed down patrilineally. This study investigated relations between major surnames and Y chromosomal polymorphism among the Japanese male population. To analyze genetic phylogeny in namesakes, the Y-single nucleotide polymorphism (SNP) plus Y-short tandem repeat (STR) approach was employed. A haplogroup based on SNPs and haplotypes at 17 STR loci were typed in 567 unrelated volunteers recruited in Kanagawa, Japan. Samples covered 27 common surnames such as Satoh and Suzuki, each name having 10-55 bearers. Significant difference was found for SNP haplogroup compositions and a multidimensional scaling plot using STR haplotypes in several surname groups. By contrast, these common surnames displayed wide diversity with phylogenetic networks, suggesting that no genetic drift event has occurred in their history. In all, 22 descent clusters were found, as judgcriteria ed by ad hoc of groups within five mutational steps in the 15 STR loci with the same haplogroup. The times of the most recent common ancestor ranged from 279 to over 2577 years. According to the approximate millennium span of Japanese surname history, descent criteria are expected to be reasonable for grouping within four step-neighbors. High heterogeneity of common surnames resembles that observed for England and Spain, but not for Ireland. Our results highlight that common Japanese surnames consist of descent clusters and many singletons, reflecting a mixture of long-term bearers and short-term bearers among the population. The genetic study of this population revealed characteristic features of Japanese surnames.

Evaluation of Y chromosomal SNP haplogrouping in the HID-Ion AmpliSeq™ Identity Panel.

The Y chromosomal haplogroup determined from single nucleotide polymorphism (SNP) combinations is a valuable genetic marker to study ancestral male lineage and ethical distribution. Next-generation sequencing has been developed for widely diverse genetics fields. For this study, we demonstrate 34 Y-SNP typing employing the Ion PGM™ system to perform haplogrouping. DNA libraries were constructed using the HID-Ion AmpliSeq™ Identity Panel. Emulsion PCR was performed, then DNA sequences were analyzed on the Ion 314 and 316 Chip Kit v2. Some difficulties became apparent during the analytic processes. No-call was reported at rs2032599 and M479 in six samples, in which the least coverage was observed at M479. A minor misreading occurred at rs2032631 and M479. A real time PCR experiment using other pairs of oligonucleotide primers showed that these events might result from the flanking sequence. Finally, Y haplogroup was determined completely for 81 unrelated males including Japanese (n=59) and Malay (n=22) subjects. The allelic divergence differed between the two populations. In comparison with the conventional Sanger method, next-generation sequencing provides a comprehensive SNP analysis with convenient procedures, but further system improvement is necessary.

Methylation analysis of circadian clock gene promoters in forensic autopsy specimens.

DNA methylation in gene promoter regions influences gene expression. Circadian clock genes play an important role in the formation of a biological clock and aberrant methylation of these genes contributes to several disorders. In this study, we examined forensic autopsy specimens to determine whether DNA methylation status in the promoter regions of nine circadian clock genes (Per1, Per2, Per3, Cry1, Cry2, Bmal1, Clock, Tim, and Ck1e) is related to a change in acquired diathesis and/or causes of death. Methylation-specific PCR and direct sequencing methods revealed that the promoters of Per1, Cry2, Bmal1, Clock, and Ck1e were unmethylated in all the forensic autopsy specimens, while the promoters of Per2, Per3, Cry1, and Tim were partially methylated. Methylation status varied between individuals and between tissues in the same patient. A detailed analysis of methylation patterns in the Cry1 promoter region revealed that the patterns also varied between individuals and the Cry1 promoter had highly methylated patterns in two cases that had been exposed to methamphetamine. These results suggest that the methylation status of clock gene promoters varies between individuals. Methamphetamine use may influence methylation in the Cry1 gene promoter region and disturb circadian rhythmicity.

Retrospective review of Japanese sudden unexpected death in infancy: the importance of metabolic autopsy and expanded newborn screening.

Sudden unexpected death in infancy is defined as sudden unexpected death occurring before 12 months of age. The common causes of sudden unexpected death in infancy are infection, cardiovascular anomaly, child abuse, and metabolic disorders. However, the many potential inherited metabolic disorders are difficult to diagnose at autopsy and may therefore be underdiagnosed as a cause of sudden unexpected death in infancy. In the present study we retrospectively reviewed 30 Japanese sudden unexpected death in infancy cases encountered between 2006 and 2009 at our institute. With postmortem blood acylcarnitine analysis and histological examination of the liver, we found two cases of long-chain fatty acid oxidation defects. Molecular analysis revealed that the one patient had a compound heterozygote for a novel mutation (p.L644S) and a disease-causing mutation (p.F383Y) in the carnitine palmitoyltransferase 2 gene. Furthermore, retrospective acylcarnitine analysis of the newborn screening card of this patient was consistent with carnitine palmitoyltransferase II deficiency. Metabolic autopsy and expanded newborn screening would be helpful for forensic scientists and pediatricians to diagnose fatty acid oxidation disorders and prevent sudden unexpected death in infancy.

Diplotype analysis of the human cardiac sodium channel regulatory region in Japanese cases of sudden death by unknown causes.

Inherited mutations in the human cardiac sodium channel (SCN5A) gene cause arrhythmogenic diseases such as tachyarrhythmia and bradyarrhythmia. Moreover, mutation subsets in the coding region impair SCN5A function, potentially leading to sudden cardiac death (SCD). In the present study, we performed diplotype analysis of the regulatory region of the SCN5A gene in Japanese people who died suddenly because of an unknown cause (sudden death group; n=70) and controls (n=112). There were no significant differences at six polymorphic loci between the groups. However, 38 diplotypes of 6-nucleotide polymorphism variants were identified. One of these diplotypes-Dip.D (CTG-TC/CCG-TC)-occurred significantly more frequently in the sudden death group than in the controls (p<0.01, OR=5.18, 95% CI: 1.38-19.45). Dip.D has two variants (T-1062C and T-847G), and while it is unclear whether these directly affect mRNA expression, a common polymorphism in this region modulates SCN5A expression in vitro. Our results thus suggest that the transcription of the SCN5A Dip.D variant may be associated with arrhythmogenic diseases that can induce sudden death.