Investigation of a DNA Profiling Method Using Only Cementum More Than 70 Years After Death.

Short tandem repeat (STR) typing is widely used not only for blood relationship identification but also for the personal identification of unidentified bodies. However, DNA is susceptible to the effects of environmental factors, consequently leading to reduced DNA yields. Therefore, to maximize the DNA yield required for identification, teeth are generally completely pulverized during DNA extraction. However, this renders subsequent testing after DNA profiling impossible. In this study, we investigated the utility of DNA profiling using only the cementum from teeth that had been left outdoors for long postmortem intervals. We analyzed 90 molars (fresh teeth) that were extracted within six months at a dental clinic and 90 molars (stale teeth) exposed outdoors for over 70 years, and following cementum extraction, the accuracy of STR profiling, optimal site for cementum collection, and minimum amount of cementum required for STR profiling were determined. The results demonstrated that the profiling accuracy of DNA extracted from cementum was comparable to that of DNA from dental pulp and dentin. Furthermore, the collection of cementum from either near the cervical line or from the root apex areas did not show significant differences in DNA profiling accuracy, indicating that securing at least 5 mg of cementum was sufficient to ensure precise DNA profiling. These findings suggest that DNA profiling using only cementum is viable even in teeth that have been subjected to a long postmortem interval.

Estimation of cadaveric age in crime scenes using Raman spectroscopy.

The number of unidentified corpses has been increasing in recent years. There is a need for an objective and readily applicable method to estimate age, which is important information for identification. In previous reports, we reported that the protein folding ratio (RPF) of skin, as measured by Raman spectroscopy using cross sections of skin samples, is highly correlated with age. In this study, we investigated the possibility of estimating age by measuring Raman spectra from the skin surface of cadavers using a portable device. The resultant intercept, slope, and root mean square error were 97.9, - 63.7 (p < 0.0001), and 11.68, respectively. We evaluated this regression formula by using 10-fold cross-validation, resulting in a coefficient of determination of 0.51. The portable Raman spectrometer may be of assistance in estimating age at death of corpses at the scene of discovery.

Comparison of Lidocaine and Mepivacaine for Variation in Regional Tissue Oxygenation in Stellate Ganglion Block: A Randomized, Double-Blind, Crossover Trial.

Introduction This prospective, randomized, double-blind, crossover trial aimed to investigate the effect of different types of local anesthetics on regional tissue blood oxygenation on the stellate ganglion block (SGB). Methods Twenty eligible patients were recruited for this study; 16 of whom were allocated to the study protocol. Participants were randomized into one of the two crossover sequences: 1% lidocaine hydrochloride or 1% mepivacaine hydrochloride, and received SGBs with 6 mL of any one of the local anesthetics, followed by a washout period of more than 24 hours, and then received SGBs by substituting the two local anesthetics. The tissue oxygenation index (TOI) in the skin near the mental foramen on the blocked side was recorded using near-infrared spectroscopy at 15 minutes after the local anesthetic injection. Results One participant did not complete the study. As such, variation in regional tissue oxygenation was compared between the 15 participants. There was no difference in the increase in regional tissue blood flow or tissue oxygenation after SGB between the patients who were given lidocaine and the ones who were given mepivacaine; however, the kinetics of the increase in regional tissue oxygenation were significantly faster with mepivacaine than with lidocaine. Conclusions Different types of local anesthetics do not affect the intensity of the increase in regional tissue oxygenation after SGB, but they do affect the kinetics of the increase. These differences in local anesthetics may affect how patients feel after receiving SGB, the duration of SGB effects, and the frequency of adverse events associated with SGB.

Influence of the amount of saliva deposition and time elapsed after deposition on bite mark analysis.

Bite mark analysis is among the most interesting research fields in forensic odontology; however, it is limited by its dependence on the employed method as well as assessor subjectivity, particularly when using morphological analysis or DNA profiling. These limitations are due to differences in DNA collected from saliva adhering to a living or inanimate body, as well as differences in exocrine fluid secretion and deposition amount among individuals. This study aimed to analyze the effectiveness of DNA profiling when there are differences in the amount of saliva adhering to a living body and when time has elapsed since deposition. Most allele peaks could be identified in 1 µl of saliva, even 9 h after saliva deposition and examination. Consistent results were obtained following saliva deposition in an individual who had engaged in up to 9 h of free activity. The results of this study demonstrate the validity and reliability of DNA profiling for bite mark analysis and are extremely important as they can demonstrate the usefulness of the little information left by a suspect on a victim's body.

A Newly Discovered Tendon Between the Genioglossus Muscle and Epiglottic Cartilage Identified by Histological Observation of the Pre-Epiglottic Space.

Epiglottic retroversion is difficult to explain anatomically. One reason is inadequate structural identification of the ligaments in the submucosal tissue anterior to the epiglottis (pre-epiglottic space, PES). Although studies have shown that tongue root movement plays a role in epiglottic retroversion, few morphological reports have investigated the attachment of the lingual muscles to the epiglottis. This study reconstructed the fiber structure of the PES by comprehensively analyzing fiber alignment in the PES focusing on the hyoepiglottic ligament, which runs between the lingual muscles and the epiglottis. Gross and microscopic observations of the submucosal structures from the tongue to the larynx of 20 cadavers (10 men, 10 women; mean age 79 years) were performed. A tendon continuing from the posterior part of the genioglossus muscle and attaching to the center of the epiglottic cartilage was identified in the midline area of the epiglottis. We named this tendon the glossoepiglottic tendon. In contrast, the hyoepiglottic ligament is found between the hyoid bone and the epiglottis and is attached from the lateral margin of the epiglottic cartilage to its base. Furthermore, the glossoepiglottic tendon consists of a high-density fiber bundle that is thicker than the hyoepiglottic ligament. These results show that the conventional hyoepiglottic ligament has a two-layer structure consisting of an upper fiber bundle connected to the genioglossus muscle and a lower fiber bundle connected to the hyoid bone. Sustained contraction of the posterior part of the genioglossus muscle therefore places the epiglottis under persistent traction, suggesting that its relaxation may cause epiglottic retroversion.

A method for estimating time since death through analysis of substances deposited on the surface of dental enamel in a body immersed in freshwater.

Estimating the elapsed time after death of a corpse immersed in freshwater is often challenging owing to body temperature changes and the degree of postmortem lividity, and the tissues used for this assessment deteriorate drastically as the postmortem period progresses. In this study, assuming a corpse was immersed in freshwater, we analyzed changes in the amount of deposits on the enamel surface of teeth since the time of immersion using an electron probe microanalyzer. We calculated a regression equation for estimating the time of immersion in water (the time after death). We considered this approach would be useful to estimate the time after death even in rotting and skeletonized corpses from which only limited information could be obtained. This study was undertaken according to a previously reported method that used a regression equation for estimating the time after death of bodies found in a seawater area.

First-in-human results of an in-body tissue architecture-induced tissue-engineered vascular graft "Biotube" for application in distal bypass for chronic limb-threatening ischemia.

Small-diameter vascular substitutes remain necessary, especially in the absence of an available autologous vein. Using a completely autologous approach termed "in-body tissue architecture," a small-diameter, long, tissue-engineered vascular graft, a "Biotube," was developed. A below-the-knee distal bypass using the Biotube as a composite with expanded polytetrafluoroethylene grafts was performed to treat a patient with chronic limb-threatening ischemia without a venous graft available. The wound on the foot had completely healed 3 months after the bypass surgery, and limb salvage and walking without claudication were achieved. At the 1-year postoperative follow-up examination, duplex ultrasound scans demonstrated graft patency without thrombus or stenosis.

Survey of dental hygienists' and dental hygienist students' attitudes towards body identification tasks of unidentified human remains.

OBJECTIVE: Dental findings are used to identify unidentified human remains. The post-mortem records, necessary for the appraisal of dental findings, are created by the dentist; however, a system for dental hygienists specializing in these tasks to utilize has not yet been considered. METHODS: This study was directed at both students at a dental hygienist school and dental hygienists. We conducted an awareness survey on their willingness to participate in these tasks, as well as the need for education regarding body identification tasks. RESULTS: The results showed that students and dental hygienists are highly interested in body identification tasks and highly motivated to participate. Numerous participants preferred to take courses regarding basic knowledge in both lecture and practical training settings to acquire the knowledge necessary for performing these tasks. CONCLUSIONS: It is necessary to consider the curriculum content and timing of lectures and practices related to body identification tasks. Participation limited to the creation of post-mortem records, except for the creation of antemortem records requiring diagnosis and treatment knowledge and the matching task of antemortem records and post-mortem records, should be considered to prevent the tasks of dental hygienists from excessively expanding. If dental hygienists train properly, intraoral findings records can be drawn up by a team consisting of 1 dentist and 1 dental hygienist. Therefore, it is expected that the use of forensic dental identification, which is a relatively quick identification method, will be further improved.

Cervical nerve roots and the dural sheath: a histological study using human fetuses near term.

We have previously reported that the thoracolumbar posterior nerve root shows a tortuous epidural course, based on studies of human fetuses near term. For comparison with the cervical nerve, examinations were conducted using frontal, sagittal and horizontal sections of cervical vertebrae from 22 fetuses at 30-38 weeks of gestation. The cervical nerve root showed a short, straight and lateral course near the zygapophysial joint. Multiple rather than single bundles of the cervical posterior root seemed to account for the majority of sensory nerve fibers innervating the upper extremity. Fasciculation of rootlets was evident near the thoracolumbar spinal cord, whereas it was seen in the dural pocket at the nerve exit from the dural sac although both sites were subdural. As in the thoracolumbar region, the nerve sheath was continuous with the dura mater and independently surrounded each of the anterior and posterior roots. Radicular arteries were few in the cervical region. In 2 of the 22 fetuses (31 weeks and 33 weeks), there was a segmental, unilateral abnormality of nerve rootlet fasciculation where the dorsal root ganglion was located lateral or peripheral to the intervertebral region. Long nerve roots running inferiorly are a necessary adaptation to the delayed and marked growth of the thoracolumbar vertebral column. In children, the cervical nerve roots are likely to be affected by movement or dislocation of the vertebrae. The segmental abnormality of the cervical nerve root may be linked to rare variations in the brachial plexus.

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.

Population history in Okinawa based on JC virus and ALDH2 genotypes.

It is widely known that people in Okinawa originated from the Jomon people, and are generally tolerant to alcohol. However, some individuals in mainland Japan lack alcohol tolerance due to a mutation in the human mitochondrial aldehyde dehydrogenase 2 (ALDH2) gene. Furthermore, the JC virus (JCV) genotype MY, which is related to the Jomon people, has not been found in Okinawa. In this study, to analyze the origin of the Okinawan people, we investigated the relationship between the JCV genotype and ALDH2 genotype. We collected 108 JCV positive samples from Okinawa. Only CY genotype JCV, and not the MY genotype, was detected. Among JCV-positive samples, a variant of ALDH2 (Glu/Lys heterozygote) was detected in 31 samples (29%) and wild-type ALDH2 (Glu/Glu homozygote) was detected in 77 samples (71%). Another variant of ALDH2 (Lys/Lys homozygote) was not detected. Among carriers of CY genotype JCV, wild-type ALDH2 was much more frequent in people living in Okinawa than in mainland Japan (P < 0.05). Our results suggested that the original inhabitants of Okinawa were people who carried MY genotype JCV and wild-type ALDH2; and that after the extinction of these original inhabitants, people who carried CY genotype JCV and wild-type ALDH2 migrated to the area. Due to the founder effect, CY genotype JCV and wild-type ALDH2 became dominant. Over a long period, many people with the variant ALDH2 migrated to Okinawa; the variant allele increased in frequency, but other JCV genotypes were eliminated.

A new method for estimating time since death by analysis of substances deposited on the surface of dental enamel in a body immersed in seawater.

The present investigation was performed with the objective of developing a method to estimate how long a corpse had been immersed in water after death (the time since death). Accurate determination of the time elapsed since death may lead to identification of the place of drowning, and therefore, serves not only as a piece of information useful for determination of the cause of death but also leads to prompt identification of the body. The results showed that diatoms attached to the surface of dental enamel increased with prolongation of immersion time in water. Further, as the immersion time increased, the quantity of O, Si, Mg, K, Al, and S detected on the surface of dental enamel increased, while the quantity of the main dental components (Ca and P) that were detected gradually decreased. Based on these results, we calculated a regression formula to estimate the immersion time. Our method is considered to be a breakthrough technique for evaluating the time since death more objectively, compared to the conventional method of determination based on the degree of decomposition of the corpse.

The relationship between the intramuscular bleeding of the tongue and cause of death.

Intramuscular bleeding of the tongue (ImBT) is occasionally found during post mortem examination. Despite its frequent reports, its cause has not yet been clarified. In this study, forensic autopsy data of 799 cadavers were examined and the relationship between ImBT and cause of death was investigated. A total of 74 cases showed ImBT (16 of 66 cases of fire fatality, 17 of 108 cases of drowning, 23 of 74 cases of asphyxiation, and 18 of 551 cases of other causes of death). The frequency of bleeding was significantly greater in cases with causes of death including fire fatality, drowning, and asphyxiation compared to those with other causes of death (p < 0.01). Among asphyxiation cases, ImBT was confirmed in two of five cases of typical hanging, three of 16 cases of atypical hanging, six of six cases of ligature strangulation, two of two cases of manual strangulation, eight of 38 cases of airway obstruction, and two of seven cases of oxygen deficiency. Among fire fatalities, the carboxyhemoglobin (CO-Hb) concentration of cases with ImBT was significantly lower than that in cases without ImBT (p < 0.01). In addition, the frequency of bleeding was significantly higher (p < 0.05) in cases where the blood cyanide concentration was 0.05 ppm or less. These observations suggested that ImBT during fire fatality occurs in a manner similar to that of ligature or manual strangulation, in which the flames cause the contraction and decrease in elasticity of the skin. Past reports indicated that bleeding frequency in cases of drowning did not significantly differ from that in cases with other causes of death. However, our cases showed a statistically higher incidence of bleeding compared to that in the other causes of death (p < 0.01). These results suggested that ImBT is a characteristic finding in cases of asphyxia and is an important evaluation for the diagnosis of death. When the relationship between ImBT and petechial hemorrhage was examined in three of the causes of death, no significant difference was observed between fire fatality and asphyxiation, but drowning was significantly different (p < 0.05). In cases without ImBT, the secretion of noradrenaline and adrenaline was significantly higher (p < 0.01). This finding suggests that it is unlikely that excessive secretion of catecholamine causes ImBT.

Utilization of bone impedance for age estimation in postmortem cases.

In the field of Forensic Medicine the number of unidentified cadavers has increased due to natural disasters and international terrorism. The age estimation is very important for identification of the victims. The degree of sagittal closure is one of such age estimation methods. However it is not widely accepted as a reliable method for age estimation. In this study, we have examined whether measuring impedance value (z-values) of the sagittal suture of the skull is related to the age in men and women and discussed the possibility to use bone impedance for age estimation. Bone impedance values increased with aging and decreased after the age of 64.5. Then we compared age estimation through the conventional visual method and the proposed bone impedance measurement technique. It is suggested that the bone impedance measuring technique may be of value to forensic science as a method of age estimation.

A new method for the estimation of age at death by using electrical impedance: A preliminary study.

Estimation of age at death is an important part of physical and forensic anthropology. The aim of this study was to investigate whether bioelectrical impedance analysis (BIA) of long bones can be used in the field of forensic medicine as a method to estimate age at death. BIA is easy to use and allows repeat measurements to be taken over time, and the equipment is inexpensive and portable. Impedance values (Z values) in 378 long bones (humeri, radii, femora, tibiae) of 40 male (233 bones) and 26 female (145 bones) autopsy cadavers were measured using two wire electrodes (connected to an alternating current device and a measuring device) inserted into the metaphyses of long bones. Computed tomography of the greater trochanter of the femur was also analyzed to evaluate bone mineral density (BMD). The results showed that Z values could be used to estimate age in males, and with an age cut-off point of 50 years in females. When estimating age at death from femur and tibia in males by using BIA, it is not necessary to consider the length of subjects. And it was thought that Z values reflected BMD. Among the long bones, the tibia is the most suitable and easy to use for Z value measurement. Z value measurement is a useful method for quantitative evaluation of age at death that can be performed quickly with inexpensive, portable equipment.

Characteristic findings on panoramic radiography and cone-beam CT to predict paresthesia after extraction of impacted third molar.

The purpose of this study was to compare findings on the relationship between impacted molar roots and the mandibular canal in panoramic and three-dimensional cone-beam CT (CBCT) images to identify those that indicated risk of postoperative paresthesia. The relationship between impacted molars and the mandibular canal was first classified using panoramic images. Only patients in whom the molar roots were either in contact with or superimposed on the canal were evaluated using CBCT. Of 466 patients examined using both panoramic and CBCT images, 280 underwent surgical extraction of an impacted molar, and 15 of these (5%) reported postoperative paresthesia. The spatial relationship between the impacted third molar root and the mandibular canal was determined by examining para-sagittal sections (lingual, buccal, inter-radicular, inferior, and combinations) obtained from the canal to the molar root and establishing the proximity of the canal to the molar root (in contact with or without loss of the cortical border and separate). The results revealed that darkening of the roots with interruption of the mandibular canal on panoramic radiographs and the inter-radicular position of the canal in CBCT images were characteristic findings indicative of risk of postoperative paresthesia. These results suggest that careful surgical intervention is required in patients with the above characteristics.

Tracing Jomon and Yayoi ancestries in Japan using ALDH2 and JC virus genotype distributions.

BACKGROUND: According to the dual structure model, the modern Japanese ethnic population consists of a mixture of the Jomon people, who have existed in Japan since at least the New Stone Age, and the Yayoi people, who migrated to western Japan from China around the year 300 bc Some reports show that the Yayoi are linked to a mutation of the aldehyde dehydrogenase 2 gene (ALDH2). Recent viral studies indicate two major groups found in the Japanese population: a group with the CY genotype JC virus (JCV) and a group with the MY genotype JCV. It is unclear whether either genotype of the JC virus is related to the Jomon or Yayoi. In this study, we attempted to detect JCV genotypes and ALDH2 mutations from the DNA of 247 Japanese urine samples to clarify the relationship between the dual structure model and the JCV genotype through ALDH2 mutation analysis and JCV genotyping. FINDINGS: The ALDH2 polymorphism among 66 JC virus-positive samples was analyzed, and it was found that the ALDH2 variant is significantly higher in the population with CY genotype JCV (51.5 %) than in the population with the MY genotype (24.2 %) (p < 0.05). CONCLUSION: From these findings, it may be inferred that the ALDH2 mutation, which is related to the Yayoi, is related to CY genotype JCV. When the Yayoi migrated to the Japanese archipelago, they brought the ALDH2 mutation as well as the CY genotype JCV.

Estimation of postmortem time based on aorta narrowing in CT imaging.

Computed tomography (CT) in forensic medicine is commonly used in determination of cause of death. Here, we show that the information provided by CT may also be of value for estimation of time after death. The relationship between abdominal aorta narrowing just below the diaphragm and time after death was examined in 140 cases. Aorta narrowing was defined using a flatness ratio, which was calculated by dividing the shorter axis of the aorta by the longer axis. Time after death was classified into 6 time periods. The flatness ratio gradually and significantly decreased with increased postmortem time, showing increased aorta narrowing with increased time after death. A further examination of aorta narrowing in 15 autopsy cases in which CT images were taken at two postmortem times gave similar findings. The results of the study suggest that aorta narrowing in postmortem CT imaging may be used to estimate time after death.

A trial of the utilization of stable isotope analysis for the estimation of the geographic origins of unidentified cadavers.

The number of unidentified cadavers is increasing worldwide and the effective methods which reveal their geographic origin are not well known. In this study, we analyzed the urine stable isotope ratio of hydrogen and oxygen collected from three locations: Chiba (Japan), Fuzhou (China), and Denpasar (Indonesia) from healthy volunteers. In addition, analysis of the effect of drinking bottled water on stable isotope ratios found in urine, and the comparison of the stable isotope ratios of urine and saliva, were conducted. Statistically significant differences in δ(2)H and δ(18)O values from the three locations were found. In this pilot study, urine δ(18)O values became increasingly similar to those of bottled drinking water during an eight-day period of drinking only bottled water. In a separate pilot study significant differences in δ(18)O, δ(13)C, and δ(15)N values from urine and saliva were found, but not in δ(2)H values. In all three studies, although the number of samples was limited, the results suggest that with further research, stable isotope analysis from urine samples might be used to identify the origins of unidentified corpses, assist in determining the length of time an individual has been in a given area and distinguish between body fluids.