Forensic application of three interstitial pneumonia markers: search for new pneumonia markers in dead bodies.

In forensic cases, detailed identification of pneumonia is important. Our objective was to statistically determine the applicability of three interstitial lung disease (ILD) markers for forensic diagnosis using serum collected from dead bodies with various postmortem intervals (PMIs). We retrospectively analyzed the levels of postmortem serum Krebs von den Lungen-6 (KL-6) and pulmonary surfactant-associated proteins A and D (SP-A and SP-D) using 221 samples obtained during forensic autopsy at our facility from 2019 to 2023. We evaluated the diagnostic efficacy of ILD markers for various pneumonias against the pathological diagnosis, and examined the assessment of the severity of ILD. When comparing the ILD group with bacterial pneumonia (BP) versus the control group, there was a significant increase in KL-6 in the ILD group. When comparing the severe ILD (SILD) group with the mild ILD (MILD) group, there was a significant increase in KL-6 and SP-D in the SILD group. The optimal cutoff values for differentiating SILD were 607.0 U/mL for KL-6, 55.5 ng/mL for SP-A, and 160.0 ng/mL for SP-D, and the sensitivity/specificity (%) of KL-6, SP-A, and SP-D for SILD were 84.1/95.2, 55.6/85.7, and 66.7/74.6, respectively. This is the first study to examine KL-6 in postmortem serum in forensic medicine. By analyzing dead bodies with various PMIs, our results confirmed statistically that postmortem serum KL-6 specifically detects ILD, postmortem serum SP-A has high sensitivity to lung injury, and postmortem serum SP-D is potentially useful in assessing the severity of ILD.

A suicide case of liquid nicotine intoxication.

A man in his 50 s, who was found vomiting and in a disturbed state when the emergency medical team arrived, then went into cardiopulmonary arrest during transport and died without responding to resuscitation. The hospital initially suspected that the death may have been caused by internal causes, but since the deceased had previously been transported to the hospital in a suicide attempt, the hospital called police regarding suspicions of unnatural death. The police investigation revealed two empty bottles of nicotine liquid for e-cigarettes in his house and a search history of "nicotine suicide" on his cellphone. In a forensic autopsy, he was found to be highly obese, and abundant fat deposits were observed in his organs. A stent was placed in the aorta, but no abnormality was found. There was no obvious stenosis or obstruction in the coronary arteries. Drug screening using liquid chromatography tandem mass spectrometry (LC-MS/MS) was performed on cardiac blood, urine, and stomach contents collected at autopsy, which revealed the presence of some medical products such as aripiprazole, nicotine, and cotinine. Further quantitative testing revealed high concentrations of nicotine in all samples. The left and right femoral venous blood concentrations were above the lethal dose, suggesting that arrhythmia or respiratory failure due to nicotine intoxication was the cause of death. With the widespread use of e-cigarettes, high concentrations of nicotine are readily available, and case reports of serious nicotine addiction are increasing. It is important to always consider addiction when conducting forensic evaluations in the medical field.

High titers of infectious SARS-CoV-2 in corpses of patients with COVID-19.

OBJECTIVES: The prolonged presence of infectious SARS-CoV-2 in deceased patients with COVID-19 has been reported. However, infectious virus titers have not been determined. Such information is important for public health, death investigation, and handling corpses. The aim of this study was to assess the level of SARS-CoV-2 infectivity in the corpses of patients with COVID-19. METHODS: We collected 11 nasopharyngeal swabs and 19 lung tissue specimens from 11 autopsy cases with COVID-19 in 2021. We then investigated the viral genomic copy number by real-time reverse transcription-polymerase chain reaction and infectious titers by cell culture and virus isolation. RESULTS: Infectious virus was present in six of 11 (55%) cases, four of 11 (36%) nasopharyngeal swabs, and nine of 19 (47%) lung specimens. The virus titers ranged from 6.00E + 01 plaque-forming units/ml to 2.09E + 06 plaque-forming units/g. In all cases in which an infectious virus was found, the time from death to discovery was within 1 day and the longest postmortem interval was 13 days. CONCLUSION: The corpses of patients with COVID-19 may have high titers of infectious virus after a long postmortem interval (up to 13 days). Therefore, appropriate infection control measures must be taken when handling corpses.

Bioprotective role of platelet-derived microvesicles in hypothermia: Insight into the differential characteristics of peripheral and splenic platelets.

BACKGROUND: Most platelets are present in peripheral blood, but some are stored in the spleen. Because the tissue environments of peripheral blood vessels and the spleen are quite distinct, the properties of platelets present in each may also differ. However, no studies have addressed this difference. We previously reported that hypothermia activates splenic platelets, but not peripheral blood platelets, whose biological significance remains unknown. In this study, we focused on platelet-derived microvesicles (PDMVs) and analyzed their biological significance connected to intrasplenic platelet activation during hypothermia. METHODS: C57Bl/6 mice were placed in an environment of -20 °C, and their rectal temperature was decreased to 15 °C to model hypothermia. Platelets and skeletal muscle tissue were collected and analyzed for their interactions. RESULTS: Transcriptomic changes between splenic and peripheral platelets were greater in hypothermic mice than in normal mice. Electron microscopy and real-time RT-PCR analysis revealed that platelets activated in the spleen by hypothermia internalized transcripts, encoding tissue repairing proteins, into PDMVs and released them into the plasma. Plasma microvesicles from hypothermic mice promoted wound healing in the mouse myoblast cell line C2C12. Skeletal muscles in hypothermic mice were damaged but recovered within 24 h after rewarming. However, splenectomy delayed recovery from skeletal muscle injury after the mice were rewarmed. CONCLUSIONS: These results indicate that PDMVs released from activated platelets in the spleen play an important role in the repair of skeletal muscle damaged by hypothermia.

Validity of dental findings for identification by postmortem computed tomography.

Postmortem computed tomography (PMCT) images help identify individuals and extract information from corpses. PMCT may substitute for a standard examination when bodies are severely damaged or when resources are limited in a mass fatality incident. In such situations, the dental information revealed by PMCT has the potential to narrow down candidates for identification further. However, the validity of the dental findings obtained from PMCT images remains unclear. In this study, we investigated the validity of dental findings on PMCT images compared to regular dental examinations as the reference standard. We routinely collected PMCT images of 148 unidentified corpses and compared the dental findings of each tooth obtained from PMCT with those of a regular dental examination. The validity of dental findings of PMCT was measured by sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). We also evaluated the accuracy of dental findings per corpse using 32 teeth as the denominator. The dental findings with high sensitivity and high specificity were a present tooth (0.96 and 0.97), a missing tooth (0.97 and 0.97), pontic (0.92 and 1.00), a dental implant (1.00 and 1.00), and a root filling (0.94 and 0.99). The mean accuracy of the dental findings per corpse in PMCT was 95.6% (standard deviation: 6.9, minimum ≤ median ≤ maximum: 65.6 ≤ 100 ≤ 100). The number of corpses with 100% accuracy in the dental findings was 81 (54.7%). The information obtained in this study highlights the potential use of PMCT during human identification in several settings with limited resources, such as the number of specialists present and the condition of the corpses.

SmartAmp method can rapidly detect SARS-CoV-2 in dead bodies.

Rapid and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in dead bodies is essential to prevent infection among those working with dead bodies. This study focused on the Smart Amplification (SmartAmp) method, which has a short examination time (approximately an hour), is simple to perform, and demonstrates high specificity and sensitivity. This method has already been used for clinical specimens; however, its effectiveness in dead bodies has not been reported. This study examined the SmartAmp method using 11 autopsies or postmortem needle biopsies performed from January to May, 2021 (of these, five cases tested positive for SARS-CoV-2 by quantitative real-time polymerase chain reaction (qRT-PCR) and six cases tested negative). Swab samples were collected from the nasopharynx, oropharynx, or anus and the SmartAmp and qRT-PCR results were compared. For the nasopharynx and oropharynx samples, the same results were obtained for both methods in all cases; however, for the anal swabs, there was one case that was positive according to qRT-PCR but negative according to the SmartAmp method. The SmartAmp method may therefore be less sensitive than qRT-PCR and results may differ in specimens with a low viral load, such as anal swabs. However, in the nasopharynx and oropharynx specimens, which are normally used for testing, the results were the same using each method, suggesting that the SmartAmp method is useful in dead bodies. In the future, the SmartAmp method may be applied not only during autopsies, but also in various situations where dead bodies are handled.

Splenic peliosis associated with spontaneous rupture and massive bleeding.

We report findings from an autopsy case who died from massive bleeding because of splenic peliosis. The case subject was an 80-year-old man who had diabetes mellitus and who was receiving hemodialysis and anticoagulant therapy. Postmortem computed tomography demonstrated massive intra-abdominal hemorrhage especially seen around the spleen. At autopsy, we found abundant hemorrhagic ascites, including a large number of clots, in the abdominal cavity. The spleen had several distinct dark red areas ranging in size from 1.5 to 2.5 cm and showed spontaneous rupture along with hematoma formation on the outside of the splenic capsule on the anterior side. From these findings, we concluded that the cause of death in this case was massive hemorrhage owing to spontaneous rupture of splenic peliosis. Although peliosis itself rarely causes death, but when it is destroyed, massive bleeding leads to death. Thus, it is necessary to know the histopathological characteristics of peliosis, in forensics.

Hypothermia causes platelet activation in the human spleen.

BACKGROUND: Accidental hypothermia results in various dysfunctions in the human body. Additionally, coagulation disorder can lead to a life-threatening condition. We previously demonstrated that platelets stored in the spleen were activated and thus triggered coagulation disorder in a mouse model of hypothermia. In the present study, we wanted to investigate if this phenomenon in mice also occurs in humans as a reaction to hypothermia. METHODS: We analyzed splenic tissue collected from 22 deceased subjects who have died from hypothermia. These samples were compared with 22 control cases not exposed to cold environment. We performed immunohistochemical staining for CD61 (a marker of all platelets) and CD62P (a marker of activated platelets). We also evaluated the morphology of platelets in the spleen with scanning electron microscopy. RESULTS: Immunohistochemical analysis revealed no significant changes in the amounts of CD61-positive platelets between the hypothermia and control cases. However, the hypothermia cases contained abundant CD62P-positive platelets compared with those of the control cases. Immunohistochemical analysis also revealed that the activated platelets formed aggregates and adhered to splenic sinusoidal endothelial cells in the hypothermia cases. However, we observed no significant fibrin formation around the activated platelets. CONCLUSIONS: Hypothermia resulted in splenic platelet activation, which may be used as a postmortem marker of hypothermia. The release of activated platelets from the spleen into to circulation upon rewarming may promote coagulation disturbances.

Preparation of dentin standard samples for age estimation based on increased aspartic acid racemization rate by heating.

Age estimation in adults based on aspartic acid racemization (AAR) provides fewer errors and higher precision than that based on bone morphology for the identification of cadavers. The technique has been established in some labs as a routine method. However, as the essential requisites for the technique, a wide age range of teeth of the same type as the target tooth must be collected for calibration for each examination. We investigated whether dentin standard samples could be prepared by increasing the AAR rate via heat. Powdered dentin was prepared from a maxillary first premolar (13 years) and heated for 0-72 h at 110 °C. The extent of AAR increased significantly with heating time and the correlation was strong (r = 0.913; p < 0.01). Similar results were found for a mandibular canine (24 years, r = 0.948; p < 0.01) and a maxillary third molar (20 years, r = 0.944; p < 0.01). We attempted to estimate the age of four maxillary first premolars of persons aged 25-58 years by using the heated samples (18 years, 12 h to 7 days). The differences between the actual and estimated ages were within ±5 years. The stability of the AAR rates in the powdered dentin during storage at 22-25 °C, 4 °C, and -30 °C was examined after 1 year and no significant changes had occurred. We were able to prepare dentin standard samples and created a calibration curve. This is a pilot study that needs to be validated before it can be used in forensic practice.

Estimation of birth year by radiocarbon dating of tooth enamel: Approach to obtaining enamel powder.

Atmospheric radiocarbon (14C) levels increased from 1955 to 1963 due to atmospheric nuclear weapon tests, and then decreased. As 14C accumulates in human tooth enamel while the enamel is being formed, 14C can be used to estimate the birth year of unidentified bodies. Measurement results of 14C content in tooth enamel using accelerator mass spectrometry vary depending on the enamel's sample site. To address this problem, a method for equalizing samples using a pulverizer was considered in this study. Regarding the tube and cone used as the pulverizer, (1) a polycarbonate tube and stainless steel cone, (2) a stainless steel tube and cone, and (3) a tungsten carbide tube and cone, were compared. In (1), the modern carbon ratio was approximately half that of the normal ratio of 100 pMC, with which accurate dating was impossible, and in (2), a high background value was obtained for IAEA-C1, which was pulverized using a reusable tube and cone. In (3), the 14C content for IAEA-C1 pulverized using reusable tube and cone, which was washed with quartz sand, was 0.31 ±â€¯0.01 pMC. This result did not show any problems regarding background value. Therefore, the use of tungsten carbide products and washing with quartz sand is recommended for 14C measurement of pulverized teeth.

Age estimation based on maturation of the medial clavicular epiphysis in a Japanese population using multidetector computed tomography.

To estimate forensic age, the Study Group of Forensic Age Diagnostics of the German Association of Forensic Medicine (AGFAD) has recommended the radiological investigation of the medial clavicular epiphysis (MCE). This study aimed to investigate the relation between the maturation of the MCE and age in a contemporary Japanese population using a standard five-stage system by Schmeling et al. and Kellinghaus substage system. A total of 207 cadavers (128 males, 79 females) of known age and sex that underwent postmortem computed tomography (CT) and subsequent forensic autopsy were obtained. The earliest appearance for stage 3c was 18.1 years in the male subjects and 18.4 years in the female subjects. Our results were consistent with those in previous studies and indicate that stage 3c is crucial for medico-legal processes in evaluating age in various countries. Similar studies on the staging of the MCE using CT images in other populations, particularly in Asians, are required.

Sex estimation based on femoral measurements using multidetector computed tomography in cadavers in modern Japan.

We aimed to reproduce the anthropometrical measurements of femoral dimensions using multi-planar reconstruction (MPR) computed tomography (CT); to assess the feasibility of sex estimation using femoral measurements through this approach, and to establish the corresponding sex estimation thresholds in the modern Japanese population. We used data on 224 cadavers (116 male and 108 female) that were subjected to postmortem CT and subsequent forensic autopsy at our department between October 2009 and July 2016. Four femoral measurements were obtained. Receiver operating characteristic (ROC) curve analysis was performed to test the overall ability of the variables in sex estimation, while the accuracy of the sex estimation was measured by the area under the ROC curve (AUC). The femoral bicondylar breadth showed the largest AUC values (left; 0.973, right; 0.974), followed by the maximum head diameter (left; 0.951, right; 0.955), and maximum femoral length (left; 0.885, right; 0.887). To the best of our knowledge, this is the first report to assess sex estimation based on femoral measurements, using MPR CT images.

Establishment of a prediction method for the mid-facial region of unknown human Mongoloid skeletal remains.

Forensic facial approximation is a technique used to estimate the antemortem facial features of unknown skeletal remains. In recent years, many researchers have reported nasal tip predictions with positive results. However, the morphological nasal features of the skull can vary widely, and it is hard to obtain accurate values using facial approximation techniques. We assumed that these variations are due to an over-dependence on the values obtained from a single distance metric factor in an anatomical area. Measurements were acquired using cephalometric radiographic images obtained from 190 Japanese individuals (90 men, aged 18-36 years and 100 women, aged 18-46 years). Soft tissue and skeletal features were traced onto acetate sheets. The orbitale (Or), porion (Po), and the Frankfurt Horizontal Plane (FHP) were plotted in addition to the rhinion (Rhi), anterior nasal spine (ANS), subnasale (Sn), prosthion, and point-A (A). From these, the following were measured: a length from rhinion to prosthion; b length from rhinion to the intersection of a line perpendicular to the anterior nasal spine; c length from the prosthion to the intersection of a line perpendicular to the anterior nasal spine; g the proportion of d/b; and f the proportion of c/b. A calculation was generated from these measurements and from proportions of a-h, and applied to the samples. An R-squared (RSQ) test and standard error (SE) were used to compare the actual and predicted values. The errors observed between the predicted and actual values were not greater than 5mm in any of the samples; 91.3% and 71.2% of predicted Sn had an error lower than 2.5 and 1.5mm respectively, from the actual. Reliable results were obtained using the method in the present study. In addition, in the process of obtaining the measurements, we found reliable proportional differences between the sexes in the piriform and axillary alveolar regions.

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

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

Sex assessment based on clavicular measurements in a modern Japanese population using multidetector computed tomography.

Accurate sex estimation from unknown human remains is crucial as an integral and foremost step toward individual identification. The aims of this study were to assess sexual dimorphism of the clavicle and develop discriminant equations for sex estimation in a modern adult Japanese population based on clavicular measurements using three-dimensional (3D) computed tomographic (CT) images. A total of 300 cadavers (150 males, 150 females) of known age and sex that underwent postmortem CT and subsequent forensic autopsy were obtained. Four measurements were performed using 3D CT reconstructed images that extracted only bones based on the CT values, as follows: the linear distances between the medial superior margins of the left and right sternal facets to the lateral points of the left and right acromial ends, and the linear distances between the medial superior margins of the left and right sternal facets to the tips of the left and right conoid tubercles. The mean values of all measurements were significantly greater for male subjects than female subjects. Univariate discriminant function analysis (DFA) provided correct sex prediction rates of 88.3-92.2%. Stepwise DFA for the left and right clavicles yielded sex classification accuracy rates of 92.2% and 91.0%, respectively. In conclusion, clavicular measurements using 3D CT images of a contemporary Japanese population may be useful for sex estimation in forensic contexts.

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

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

Stature estimation in a contemporary Japanese population based on clavicular measurements using multidetector computed tomography.

The aims of this study was to assess the correlation between stature and clavicular measurements in a contemporary Japanese population using three-dimensional (3D) computed tomographic (CT) images, and to establish regression equations for predicting stature. A total of 249 cadavers (131 males, 118 females) underwent postmortem CT scanning and subsequent forensic autopsy between October 2011 and May 2016 in our department. Four clavicular variables (linear distances between the superior margins of the left and right sternal facets to the anterior points of the left and right acromial ends and between the superior margins of the left and right sternal facets to the left and right conoid tubercles) were measured using 3D CT reconstructed images that extracted only bone data. The correlations between stature and each of the clavicular measurements were assessed with Pearson product-moment correlation coefficients. These clavicular measurements correlated significantly with stature in both sexes. The lowest standard error of estimation value in all, male, and female subjects was 3.62cm (r2=0.836), 3.55cm (r2=0.566), and 3.43cm (r2=0.663), respectively. In conclusion, clavicular measurements obtained from 3D CT images may be useful for stature estimation of Japanese individuals, particularly in cases where better predictors, such as long bones, are not available.

Incidence of three-rooted mandibular first molars among contemporary Japanese individuals determined using multidetector computed tomography.

PURPOSE: The purpose of this study was to determine the incidence of three-rooted mandibular first molars in a contemporary Japanese population using multidetector computed tomography (MDCT) and examine whether this characteristic root form may be useful for identification purposes. METHODS: Prior to their forensic autopsies, we obtained MDCT scans of 365 cadavers (255 males, 110 females) with mandibular first molars on both sides. Altogether, 730 mandibular first molars were examined morphologically on reconstructed CT images, and the incidence of three-rooted molars was recorded. The results were analyzed statistically to determine sex and left-right differences using the chi-square test. RESULTS: In all, 189 (25.9%) of the 730 mandibular first molars had three roots: 79 (31.0%) in the 255 males and 35 (31.8%) in the 110 females. No statistically significant difference was found between the sexes. In the 39 individuals who had unilateral three-rooted mandibular first molars, 7 (17.9%) were on the left side and 32 (82.1%) were on the right side, indicating a statistically significant predominance on the right side. CONCLUSIONS: The incidence of three-rooted mandibular first molars in contemporary Japanese individuals was 25.9%, with no statistically significant sex difference, but in the case of unilateral three-rooted teeth, with a statistically significant predominance on the right side. Our study found that Japanese and northeastern Asians have a high incidence of the three-rooted mandibular first molars among individuals of Mongolian origin; this finding may be a useful screening tool for identification of unknown individuals.

A rare autopsy case of traumatic rhabdomyolysis associated with intermittent assault.

Traumatic rhabdomyolysis generally occurs after severe blunt trauma and is acute in onset, associated with severe disease, and potentially lethal. Accordingly, diagnosis of traumatic rhabdomyolysis in patients without massive subcutaneous or intramuscular hemorrhage is difficult, especially in the postmortem period, which is limited in terms of the availability of biochemical examination tools and accurate history of illness. To the best of our knowledge, there are no previous reports of death from traumatic rhabdomyolysis among individuals who did not pursue medical consultation. A previously healthy man in his early sixties had been punched and kicked several times in the previous 2months, but he had not gone to a hospital. He suddenly lost consciousness at his workplace approximately 5days after the most recent assault, and cardiopulmonary arrest occurred when the emergency service arrived. He died the same day, and a medicolegal autopsy was performed. Although several sites of minor subcutaneous and muscle hemorrhage were observed, the cause of death was unclear upon macroscopic assessment. Immunohistochemical staining revealed acute renal failure caused by rhabdomyolysis. We herein report a rare case of fatal traumatic rhabdomyolysis, seemingly associated with minor and apparently nonlethal muscle injury.

Sexual determination based on multidetector computed tomographic measurements of the second cervical vertebra in a contemporary Japanese population.

Accurate sex estimation is important in forensic investigation to determine the identity of unknown individuals. The aim of this study was to investigate the accuracy of sex assessment based on measurements of the second cervical vertebra (C2) using computed tomographic (CT) images in a Japanese population and to develop discriminant function formulae. The data were collected from 224 Japanese cadavers (112 male subjects, 112 female subjects) on which postmortem CT scanning and subsequent forensic autopsy were performed. Nine CT measurements of the C2 were performed for CT images of each subject. The measurements were assessed using descriptive statistics and discriminant function analyses (DFA). All of the measurements demonstrated significant sexual dimorphism. Multiple DFA with stepwise variable selection resulted in multivariable models; a five-variable model reached an accuracy rate of 92.9%. Our results suggest that metric analysis based on CT images of the C2 can accurately determine the sex from the human skeletal remains in a contemporary Japanese population and may be useful for sex estimation in forensic anthropology.