Artificial intelligence-based forensic sex determination of East Asian cadavers from skull morphology.

Identification of unknown cadavers is an important task for forensic scientists. Forensic scientists attempt to identify skeletal remains based on factors including age, sex, and dental treatment remains. Forensic scientists commonly consider skull or pelvic shape to evaluate the sex; however, these evaluations require sufficient experience and knowledge and lack objectivity and reproducibility. To ensure objectivity and reproducibility for sex evaluation, we applied a gated attention-based multiple-instance learning model to three-dimensional (3D) skull images reconstructed from postmortem head computed tomography scans. We preprocessed the images, trained with 864 training data, validated the model with 124 validation data, and evaluated the performance of our model in terms of accuracy with 246 test data. Furthermore, three forensic scientists evaluated the 3D skull images, and their performances were compared with those of the model. Our model showed an accuracy of 0.93, which was higher than that of the forensic scientists. Our model primarily focused on the entire skull owing to visualization but focused less on the areas often investigated by forensic scientists. In summary, our model may serve as a supportive tool to identify cadaver sex based on skull shape. Further studies are required to improve the model's performance.

A suicide case by hanging without fixing the end of the rope.

Hanging is a method of death in which one end of a cord is tied around the neck and the other end is tied to an unmovable object and hung down, compressing the neck with weight. We have identified a rare case in which death was accomplished without tying one end of the rope. The individual was successfully hanged by the frictional force between the upward rope and the downward rope. The police concluded the case as a suicide.

Deep Neural Networks-Based Age Estimation of Cadavers Using CT Imaging of Vertebrae.

Although age estimation upon death is important in the identification of unknown cadavers for forensic scientists, to the best of our knowledge, no study has examined the utility of deep neural network (DNN) models for age estimation among cadavers. We performed a postmortem computed tomography (CT) examination of 1000 and 500 male and female cadavers, respectively. These CT slices were converted into 3-dimensional images, and only the thoracolumbar region was extracted. Eighty percent of them were categorized as training datasets and the others as test datasets for both sexes. We fine-tuned the ResNet152 models using the training datasets. We conducted 4-fold cross-validation, and the mean absolute error (MAE) of the test datasets was calculated using the ensemble learning of four ResNet152 models. Consequently, the MAE of the male and female models was 7.25 and 7.16, respectively. Our study shows that DNN models can be useful tools in the field of forensic medicine.

Application of Teeth in Toxicological Analysis of Decomposed Cadavers Using a Carbamazepine-Administered Rat Model.

In a regular autopsy, blood and organs are used to quantify drug and toxicant concentrations; however, specimens such as blood cannot be collected from highly decomposed corpses, making the quantification of drug and toxicants impossible. This study aimed to estimate the blood carbamazepine (CBZ) concentration from teeth, a part of the human body that is best preserved after death. We sampled teeth and blood of rats administered CBZ. The correlation between the tooth and serum CBZ concentrations was analyzed. Rats were euthanized after CBZ administration and kept at 22 °C for 0 to 15 days before sampling the teeth and measuring the CBZ concentration. Undecalcified, fresh, frozen sections of rat teeth were prepared, and CBZ localization was evaluated. CBZ concentrations in both teeth and cardiac blood peaked at 60 min after administration and increased in a dose-dependent manner. CBZ concentration in teeth did not substantially change after death, with high CBZ distribution being observed in the pulp cavity. The tooth and serum CBZ concentrations were highly correlated, suggesting that the measurement of toxicant concentration in sampled teeth would allow for the estimation of blood toxicant concentration in highly decomposed corpses.

Estimating the age at death for forensic cases using quantitative computed tomography.

Estimation of the age at death is an important task for forensic scientists. Although the correlation between age and bone mineral density is already known, including for cadavers, to our knowledge, there are no published studies on age estimation with quantitative computed tomography. Quantitative computed tomography can be used to measure bone mineral density based on the mean computed tomography value of the cancellous bone. As this value cannot be calculated in putrefied cases, we modified quantitative computed tomography to calculate the bone mineral density from regions of the bone with mean computed tomography values of 50-350 Hounsfield units. We aimed to examine whether this method could be used for age estimation. We examined 171 male and 106 female cadavers, some of which were putrefied. We performed univariate linear regression analysis for age at death and bone mineral density. The resultant intercept, slope, and root mean square error were 91.3, - 0.20 (p < 0.0001), and 11.4, respectively, for male cadavers, and 96.1, - 0.23 (p < 0.0001), and 11.0, respectively, for female cadavers. We evaluated this regression formula by using 10-fold cross-validation, resulting in a coefficient of determination of 0.33 for male cadavers and 0.42 for female cadavers. The modified quantitative computed tomography method may be of assistance in estimating age at death, even in putrefied cases.

Sudden death of a 2-year-old child due to alpha-ketoadipic aciduria.

Alpha-ketoadipic acid is one of the metabolic intermediates of lysine and tryptophan, and it is known as the biochemical hallmark of alpha-ketoadipic aciduria (α-KA). α-KA is a rare autosomal recessive disorder. Its pathophysiology is reduced alpha-ketoadipic acid dehydrogenase activity, and that makes it difficult to metabolize lysine and tryptophan. The symptoms of this disease are multiple, e.g., psychomotor retardation, epilepsy, and ataxia, and it can even be asymptomatic. We present a case of sudden death in a 2-year-old boy with alpha-ketoadipic aciduria. Postmortem computed tomography (CT) and autopsy were performed to elucidate the cause of death. No obvious lesions could be identified except for a marked fatty liver. Urinalysis showed elevated excretion of α-ketoadipic acid.

The coronary artery calcification severity on postmortem CT could be a clue for the diagnosis of sudden cardiac death.

About two-thirds of sudden deaths are sudden cardiac deaths (SCD), and ischemic heart disease (IHD) accounts for 60% of these. Although an autopsy needs to be performed to prove SCD, the forensic autopsy rate is very low in Japan. To diagnose the cause of death, postmortem computed tomography (PMCT) is often performed. Because coronary artery calcification (CAC) is a risk factor in cardiac diseases such as IHD and its severity can be evaluated with CT, we examined its ability to diagnose SCD. We collected 104 autopsy cases with CT scans. On the basis of the autopsy report, we separated the cases into two groups: SCD suspected as the cause of death and SCD not suspected. We calculated each CAC severity with the Agatston score from the CT images. Cases with Agatston scores of more than 400 were labeled as severe. The relationship between SCD and CAC severity was confirmed with Fisher's exact test (p < 0.05). The sensitivity and specificity of CAC severity for SCD were 20.3% and 97.5%, respectively, and the positive likelihood ratio was 8.1. Severe CAC can increase the probability of SCD. In cases in which only PMCT can be performed, this finding can be helpful for diagnosing SCD.

Utilization of oral check-up data of autopsy cases.

The maintenance of oral health contributes to the overall health of an individual. Similar to the Adult Dental Health Survey in the UK, the Survey of Dental Diseases (SDD) is being conducted in Japan. However, our routine oral examination of autopsy cases showed that the oral cavities of the autopsied bodies were unhealthy as compared to the SDD data. Therefore, this study sought to understand the actual condition of the oral cavity and to clarify the factors related to the degradation of the oral status. This was achieved by comparing the data of the SDD subjects and autopsy cases in terms of the average number of teeth lost, the average total number of caries (number of decayed, missing, or filled teeth), percentage of people with untreated teeth, and 80-20 achievement rate (percentage of people aged 80 years with 20 or more teeth). Furthermore, this study investigated the relationship between the average number of missing teeth and smoking, psychiatric disorders, and body mass index (BMI) in autopsy cases. The oral condition of all autopsy cases was worse than that of SDD subjects. The 80-20 achievement rate was observed in more than half of the SDD subjects and only in less than 20% of the autopsy cases. Moreover, the average number of missing teeth was not significantly different depending on smoking status, BMI, and presence of psychiatric disorders in the autopsy cases. Unlike the SDD volunteers, there was no bias related to the inclusion of autopsy cases. Therefore, by analysing the data of both autopsy cases and SDD subjects, it may be possible to evaluate the oral condition more realistically. This could contribute to the realisation of improved health and quality of life.

Rapid infusion of excessive phenytoin: A newborn autopsy case.

The patient was a two-day-old female infant. The patient's mother was a primigravid in her 20 s who developed premature abruption of the normal placenta on the first day of the 33rd week of gestation. The infant was born by emergency cesarean section with severe neonatal asphyxia with a birth weight of 1928 g. Spontaneous circulation was returned 11 min after birth. The infant was treated under mechanical ventilation in the neonatal intensive care unit, and phenobarbital was administered for repeated seizures. On day 2, spontaneous respiration was observed; however, the patient developed seizures repeatedly. The dose of phenobarbital reached the maximum and was switched to midazolam. In the early morning of day 3, while midazolam was administered up to the maximum dose, the infant developed status epilepticus, and the anticonvulsant drug was changed to phenytoin. Due to a calculation error, the intravenous administration of phenytoin was started at 400 mg/30 min, which is 10-fold of the normal dose. Six minutes later, after 80 mg was administered, the administration was stopped due to a drop in blood pressure; however, the infant died of cardiac arrest. An autopsy, which was performed approximately 25 h after death, revealed the blood phenytoin concentration in the heart was 63.85 µg/mL. The cause of death was determined to be acute phenytoin toxicity. This is the first fatal case reported of the blood concentration of phenytoin caused by rapid intravenous administration.

Relationship between serum prostate-specific antigen and age in cadavers.

OBJECTIVES: An increase in number of unidentified cadavers is a growing problem. To identify these cadavers, a simple objective method is required to estimate cadaveric age. We examined the correlations between postmortem serum prostate-specific antigen levels and cadaveric age to determine whether serum prostate-specific antigen levels can be used in age estimation of unidentified cadavers. METHODS: Total serum prostate-specific antigen was measured in 140 male autopsy cases aged from 0 to 94 years. RESULTS: The serum prostate-specific antigen levels of cadavers correlated with age at death to the same degree as with the age of living individuals (r = 0.393, P < 0.01). Prostate-specific antigen levels also correlated with prostate weight, but not with psoas muscle index and body mass index. Cause of death did not influence postmortem serum prostate-specific antigen levels. CONCLUSION: Age estimation based on prostate-specific antigen provides a simple, objective, and rapid method to determine age at death estimation of cadavers, and is expected to greatly contribute to the identification of cadavers.

Estimation of Cadaveric Age by Ultrasonography.

(1) Background: While decreasing bone mineral density (BMD) with age in living people has been well documented, a correlation between age and bone mineral density in deceased people has only been reported in a few studies. A correlation between age and BMD in deceased people was investigated as an objective tool for age estimation of unidentified remains. (2) Methods: The Bone Area Ratio (BAR) was measured in 402 autopsy cases (143 females and 259 males over the age of 20). (3) Results: The correlation coefficient in the females was r = -0.5476, and the correlation coefficient in the males was r = -0.2166, indicating a stronger correlation in females than in males. A comparison of the BAR values in the deceased females for each age group with that in live females found no significant differences in the BAR values. BAR values in the deceased were similar to in live individuals, and this did not change with duration of the postmortem interval. (4) Conclusions: Measuring the BAR value based on bone mass using ultrasonic waves is rapid and easy, even for those lacking forensic training, and may be used to estimate the age of an individual and the likelihood of fracture due to trauma.

Autoerotic asphyxia using a plastic bag loosely covering the head over a gas mask.

Autoerotic death is defined as the accidental death of an individual while masturbating, caused by a device or material used to enhance the sexual response. Here we report an autopsy case of autoerotic death and review the literature. A healthy, single, 33-year-old Japanese male was found dead in his room in a prone position. He was wearing a used gas mask and a plastic bag over his head; the opening of that bag was loosely secured around the neck with a belt. He had no underwear beneath his upper and lower work clothes and the zipper of his pants was open. Adult magazines and DVDs featuring male subjects and other adult toys were found in his room. Images of people wearing gas masks and rubber suits, as well as of individuals whose whole bodies were tied with ropes, were discovered on his personal computer. Records of purchasing full-body tights were also found. Lesions and injuries that could have caused his death were not found in the autopsy or in any of the various examinations. The cause of death was determined to be suffocation assumed to be caused by the plastic bag covering the mask's ventilation. Suicide and homicide were ruled out based on the police investigation of the scene of the victim's death and the attendant circumstances and environment. The death was thus determined to be an accident. We see no major differences in the feature of autoerotic death between Japanese cases and those in the Western countries.

Comparison of postmortem butane distribution between two fatal butane poisoning cases.

Fatal accidents during butane abuse frequently occur in Japan and in many countries around the world. Although analytical data about butane concentration in postmortem samples is being accumulated, when using the data to determine the cause of death, careful interpretation is required because the gas is easily diffused. Two fatal butane poisoning cases were encountered, and butane quantification of autopsy samples obtained from left and right heart blood, femoral blood, kidney, liver, lung, brain and fatty tissues was performed. In both cases, butane concentration in the left heart blood was lower than in the right heart blood or the femoral blood, despite gas inhalation. These findings may indicate that the deceased individuals interrupted gas inhalation and inhaled room air immediately before their death, therefore ruling out asphyxia due to anoxia as the mechanism of death. Case 1, which was suspected to be a not acute death, showed a very high butane concentration ratio of fatty tissues to femoral blood of over 70. Case 2 was considered an acute death, and the butane concentration ratio of fatty tissues to femoral blood was 8.2. These results are consistent with previously reported findings showing that much higher ratios of fatty tissues to blood are compatible with long survival time. In conclusion, the comparison of butane concentration among different samples, including left heart blood versus right heart blood and fatty tissues versus blood, is useful when interpreting the result of postmortem butane analysis to examine the mechanism of death and survival time.

Infant fatality case with excessive chylous ascites.

An 11-month-old boy with marked abdominal distension was found dead in the prone position at home. Since there were many bruises in the non-protruding regions of the head, face, and abdomen, a medicolegal autopsy was performed the following day. The boy was smaller than average (height: 68.5 cm; weight: 7.8 kg); his extremities were thin; and his abdomen was remarkably bulging. Chylous ascites (1600 mL) was observed in the peritoneal cavity and chylous pleural effusion (left: 5 mL; right: 10 mL) in the thoracic cavity. A fibrous induration, approximately 2.0 × 1.5 cm in size, was observed in the root of the small bowel mesentery. Congenital chylothorax and chylous ascites were suspected. However, the remarkably withered thymus and an old injury in the superior labial frenulum suggested that the chylous ascites may have been further deteriorated by injuries sustained during physical abuse. Examination suggested that the death was sudden. Thus, we inferred that the cause of death was circulatory and respiratory failure due to excessive chylous ascites. Among the reported cases of chylous ascites in pediatric patients, some patients experiencing abuse were identified on the basis of their chief complaints of vomiting or abdominal distension. Medical and child welfare staff should be made aware of this information.

Comparison between Temperature Gradient Gel Electrophoresis of Bacterial 16S rDNA and Diatom Test for Diagnosis of Drowning.

When a body is discovered in water, it is difficult to conclude whether the cause of death was drowning, even today. Although diatom testing by the digestive method is classical, we hypothesized that aquatic bacteria, as well as diatoms, might be detected in drowned bodies, and conducted temperature gradient gel electrophoresis (TGGE)-targeting 16S rDNA. DNA was extracted from the site water, and from heart blood and liver samples from 27 bodies concluded as drowning deaths by autopsy and subjected to TGGE after amplification of 16S rDNA by polymerase chain reaction. We observed whether the feature point of each 16S rDNA from the site water and blood or liver samples matched. Considerably higher correspondence was observed in drowned bodies, and the rate was higher than that achieved with the digestive method. Moreover, TGGE is safer than the digestive method. Our study suggests that this method can aid diagnosis of drowning.

Vibrational monitor of early demineralization in tooth enamel after in vitro exposure to phosphoridic liquid.

The Raman spectroscopic method has been applied to quantitatively assess the in vitro degree of demineralization in healthy human teeth. Based on previous evaluations of Raman selection rules (empowered by an orientation distribution function (ODF) statistical algorithm) and on a newly proposed analysis of phonon density of states (PDOS) for selected vibrational modes of the hexagonal structure of hydroxyapatite, a molecular-scale evaluation of the demineralization process upon in vitro exposure to a highly acidic beverage (i.e., CocaCola™ Classic, pH=2.5) could be obtained. The Raman method proved quite sensitive and spectroscopic features could be directly related to an increase in off-stoichiometry of the enamel surface structure since the very early stage of the demineralization process (i.e., when yet invisible to other conventional analytical techniques). The proposed Raman spectroscopic algorithm might possess some generality for caries risk assessment, allowing a prompt non-contact diagnostic practice in dentistry.

Generation of Directly Converted Human Osteoblasts That Are Free of Exogenous Gene and Xenogenic Protein.

Generation of osteoblasts from human somatic cells may be applicable in an effective transplantation therapy against bone diseases. Recently we established a procedure to directly convert human fibroblasts into osteoblasts by transducing some transcription factor genes via retroviral vectors. However, retroviral vector-mediated transduction may potentially cause tumor formation from the infected cells, thus a non-viral gene transfection method may be more preferable for preparation of osteoblasts to be used for transplantation therapy. Here, we constructed a plasmid vector encoding Oct4, Osterix, and L-Myc that were an appropriate combination of transcription factors for this purpose. Osteoblast-like phenotypes including high alkaline phosphatase (ALP) activity, bone matrix production and osteoblast-specific gene expression were induced in normal human fibroblasts that were transfected with the plasmid followed by culturing in osteogenic medium. The plasmid-driven directly converted osteoblasts (p-dOBs) were obtained even in the absence of a xenogenic protein. The plasmid vector sequence had fallen out of the p-dOBs. The cells formed deposition of calcified bodies in situ after transplantation into mice. These results strongly suggest that p-dOBs can be put into practical use for a novel cell-based therapy against bone diseases. J. Cell. Biochem. 117: 2538-2545, 2016. © 2016 Wiley Periodicals, Inc.

Biomechanical force induces the growth factor production in human periodontal ligament-derived cells.

Although many reports have been published on the functional roles of periodontal ligament (PDL) cells, the mechanisms involved in the maintenance and homeostasis of PDL have not been determined. We investigated the effects of biomechanical force on growth factor production, phosphorylation of MAPKs, and intracellular transduction pathways for growth factor production in human periodontal ligament (hPDL) cells using MAPK inhibitors. hPDL cells were exposed to mechanical force (6 MPa) using a hydrostatic pressure apparatus. The levels of growth factor mRNA and protein were examined by real-time RT-PCR and ELISA. The phosphorylation of MAPKs was measured using BD™ CBA Flex Set. In addition, MAPKs inhibitors were used to identify specific signal transduction pathways. Application of biomechanical force (equivalent to occlusal force) increased the synthesis of VEGF-A, FGF-2, and NGF. The application of biomechanical force increased the expression levels of phosphorylated ERK and p38, but not of JNK. Furthermore, the levels of VEGF-A and NGF expression were suppressed by ERK or p38 inhibitor. The growth factors induced by biomechanical force may play a role in the mechanisms of homeostasis of PDL.

Electrical stimulation with periodic alternating intervals stimulates neuronal cells to produce neurotrophins and cytokines through activation of mitogen-activated protein kinase pathways.

Peripheral neuropathy is a representative complication of dental surgery. Electrical therapy, based on electrical stimulation with periodic alternating intervals (ES-PAI), may promote nerve regeneration after peripheral nerve injury in a non-invasive manner, potentially providing an effective therapy for neuropathy. This study aimed to analyze the molecular mechanisms underlying the nerve recovery stimulated by ES-PAI. In brief, ES-PAI was applied to a neuronal cell line, Neuro2A, at various intensities using the pulse generator apparatus, FREUDE. Cell viability, neurotrophin mRNA expression, and cytokine production were examined using a tetrazolium-based assay, real-time RT-PCR, and ELISA, respectively. Mitogen-activated protein kinase (MAPK) signaling was assessed using flow cytometry. It was found that ES-PAI increased the viability of cells and elevated expression of nerve growth factor (NGF) and neurotrophin-3 (NT-3); ESPAI also augmented vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) expression, which was restored by addition of p38 inhibitors. Phosphorylation of p38 and extracellular signal-regulated kinase 1/2 (ERK-1/2) was augmented by ES-PAI. Hence, ES-PAI may ameliorate peripheral neuropathy by promoting neuronal cell proliferation and production of neurogenic factors by activating p38 and ERK-1/2 pathways.

Vibrational algorithms for quantitative crystallographic analyses of hydroxyapatite-based biomaterials: II, application to decayed human teeth.

A systematic investigation, based on highly spectrally resolved Raman spectroscopy, was undertaken to research the efficacy of vibrational assessments in locating chemical and crystallographic fingerprints for the characterization of dental caries and the early detection of non-cavitated carious lesions. Raman results published by other authors have indicated possible approaches for this method. However, they conspicuously lacked physical insight at the molecular scale and, thus, the rigor necessary to prove the efficacy of this spectroscopy method. After solving basic physical challenges in a companion paper, we apply them here in the form of newly developed Raman algorithms for practical dental research. Relevant differences in mineral crystallite (average) orientation and texture distribution were revealed for diseased enamel at different stages compared with healthy mineralized enamel. Clear spectroscopy features could be directly translated in terms of a rigorous and quantitative classification of crystallography and chemical characteristics of diseased enamel structures. The Raman procedure enabled us to trace back otherwise invisible characteristics in early caries, in the translucent zone (i.e., the advancing front of the disease) and in the body of lesion of cavitated caries.