Survey of dental diseases in forensic autopsy cases.

Forensic odontology plays an important role in the age estimation of a corpse. However, some cases exhibit a large margin of error when using conventional morphological methods. The final purpose of this study was to survey dental diseases to find the factors that result in errors in age estimation. First, we surveyed dental status related to dental caries in forensic autopsy cases to examine their characteristics. We investigated 335 forensic autopsy cases with accurate age and gender information available from autopsy and police reports. We assessed the number of teeth with caries, including the number of present teeth, missing teeth, sound teeth, treated teeth, untreated teeth, and the total number of carious teeth (DMF teeth: decayed, missing, and filled teeth).Then, each mean number of teeth from autopsy cases was compared with that from the results of a control survey of dental disease performed by Akita Prefecture. The autopsy cases, mainly in their 50's, showed a significantly higher incidence of DMF teeth, missing teeth, and untreated teeth. They also showed lower incidence of treated teeth. The more severe carious status in the autopsy cases suggested that they did not regularly visit dental clinics while they were alive. We will continue to analyze their past physical disease histories, the causes of death, and their antemortem lifestyles to find the factors that lead to incorrect age estimation.

Indication and limitations of using palatal rugae for personal identification in edentulous cases.

To explore the availability and the limitations of using the palatal rugae pattern in forensic practice for personal identification in edentulous cases, we set up an experiment, which involved 48 patients who had both old and new complete dentures and we observed their rugae. First, we made 48 pairs of maxillary casts from their old and new dentures and a further 50 maxillary casts from complete dentures at random, to use as variables. All the initial impressions taken from the mucosal surfaces of complete dentures were made from alginate impression materials, and the maxillary casts were made from hard dental plaster. Secondly, all 146 casts were trimmed so that all the areas except for the rugae area were removed. Subsequently, 50 examiners were given the 48 casts from the old dentures and were then asked to compare them with the other 98 casts for possible matches. The case numbers, which matched correctly, were recorded. The median percentage of correct matches among the 50 examiners was 94%, despite variations in their experience with forensic identification, and this accuracy does not differ significantly from that in dentate cases, as described in previous publications. The median percentage of correct matches among the 48 cases was 90%. Analyzing the incidence of obtaining a correct match in each case revealed that there were three major misleading shapes that could give rise to a low rate of correct matches; (1) severely low and poorly demarcated eminences of rugae, (2) change of palatal height, and (3) non-complex rugae pattern. These features are mainly due to the shape of the edentulous palate itself and rarely due to the dentures, and could lead to difficulties in finding unique points for use in matching rugae patterns. The results suggest that an appropriate selection of cases, taking into consideration the above misleading shapes, may establish an increased rate of accuracy for identification with this method, thereby bringing the percentage of correct matches closer to 100% in edentulous cases, which is also the percentage of correct matches previously reported in dentate cases.

Population data of six STR loci using hexaplex PCR amplification and typing system, "Midi-6" in four regional populations in Japan.

The genetic differences of the allele frequency distributions for six STR loci (D20S480, D6S2439, D6S1056, D9S1118, D4S2639, and D17S1290) among regions in Japan were examined using our recently designed hexaplex amplification and typing system, "Midi-6" newly named, to construct a database in the Japanese population. Genotypes at six loci were analyzed in 198, 200, 175, and 196 individuals from the area of Akita, Nagoya, Oita, and Okinawa, respectively, in Japan. The allele frequency distributions were significantly different (p<0.05) at from one to five loci among the four populations when compared pairwise. Significant differences were also observed at two or three loci between Oita- or Okinawa-Japanese and the "pooled" population (n=769), respectively. However, since F(ST) (theta) values were extremely low (<0.05), ranging from 0.0020 to 0.0118 for six loci, genetic differentiation within the pooled Japanese population was negligible. Therefore, it suggested that the data of the allele frequencies at six loci in the pooled population would be employed as the base of calculation for statistical probabilities.

Pathological demonstration of rapid involvement into the subcutaneous tissue in a case of fatal hydrofluoric acid burns.

We report an autopsy case of a man who suffered accidental chemical burns following exposure to 60% hydrofluoric acid. The extent of the burns covered about 30% of his body surface, and cardiopulmonary arrest occurred about 30min after the exposure. At autopsy, the skin of the affected area showed greenish gray or black coloring with thin circumferential erythema, and this discoloration extended as far as the periosteum of the skull. However, such discoloration was not found on the mucosa of the airway or the gastrointestinal tract. Microscopically, severe liquefactive necrosis was already evident on the skin. Elastic fibers within the dermis were completely lost, and the entire wall of large vessels within the subcutaneous layer was already severely affected. Blood analysis in the emergency room showed hypocalcemia, and the levels of fluoride ions in the postmortem blood and urine showed extremely high values. However, fewer fluoride ions were detected from the lung tissue. The present case suggests that the hydrofluoric acid had immediately penetrated down into the deep layer of the skin, thereby involving the large vessels present within the subcutaneous layer. These pathological findings of the skin seen in the present case explain the mechanism behind the rapid dissemination of fluoride ions which entered the bloodstream from damaged arteries, resulting in the development of acute toxicity.

Relationship between cardiopulmonary resuscitation and injuries of the cardiac conduction system: pathological features and pathogenesis of such injuries.

OBJECTIVE: The purpose of this study was to assess the incidence, features, and pathogenesis of microscopic injuries to the cardiac conduction system caused by cardiopulmonary resuscitation. DESIGN: : Prospective study. SETTING: Autopsy unit of the university. PATIENTS: Victims who had been transferred to the emergency room due to cardiac arrest arising from nontraumatic cause plus age-matched control patients. INTERVENTIONS: None. MEASUREMENT AND MAIN RESULTS: The cardiac conduction system of 80 hearts without gross injury from patients who had received cardiopulmonary resuscitation due to nontraumatic causes was examined. Of these 80 patients, seven (9%) showed fresh injuries, including a lesion that had gone unreported in the previous literature. Localized hemorrhage without inflammatory reaction was evident in six of these patients. Three of the six patients showed hemorrhage in the sinoatrial node, whereas the other three patients showed hemorrhage in the atrioventricular conduction system. The remaining one patient showed localized dissection of the atrioventricular node artery with the appearance of red blood cells in the false lumen. There was no significant difference with regard to age, gender, cause of cardiopulmonary arrest, whether victim had received electrical shock treatment, whether victim had received anticoagulants, and the duration of cardiopulmonary resuscitation between the seven patients with fresh injuries and the other 73 patients. Fracture of the sternum or rib was found in only one of the seven patients but in 14 of the 73 patients. No pathologic lesions were found in the 30 control patients who did not receive cardiopulmonary resuscitation. CONCLUSIONS: It can be presumed that injuries to the conduction system do occur in limited regions during cardiopulmonary resuscitation. Minute differences in the location of the cardiac silhouette or cardiac conduction system also need to be considered, rather than just the severity of force to the anterior chest, when determining the pathogenesis of these injuries.

Simple and sensitive determination of free and total morphine in human liver and kidney using gas chromatography-mass spectrometry.

We developed a reliable, simple and sensitive method to determine free and total morphine in human liver and kidney, using gas chromatography-mass spectrometry (GC-MS). Free morphine or total morphine obtained by acid hydrolysis from 0.2g tissue sample was extracted using an Extrelut NT column with an internal standard, dihydrocodeine, followed by trimethylsilylation. The derivatized extract was submitted to GC-MS analysis of EI-SIM mode. The calibration curves of morphine in both liver and kidney samples were linear in the concentration range from 0.005 to 5 microg/g. The lower limits of detection of morphine were 0.005 microg/g. This method proved successful when we determined free and total morphine in liver and kidney obtained from an autopsied man who was mis-ingested morphine compound in the hospital, which resulted in the cause of death being morphine intoxication.

Fatal hemopericardium caused by retrograde acute abdominal aortic dissection: an autopsy report and morphological consideration.

A case of acute aortic dissection in which entry is formed at the abdominal aorta is unusual. In addition, an autopsy case of cardiac tamponade caused by spontaneous acute retrograde abdominal aortic dissection is extremely rare and such a report has never previously been published to our knowledge. We herein present an autopsy case of sudden unexpected death due to retrograde extension that originated from the abdominal aorta, together with our considerations regarding the etiology and the unfortunate course of the victim.

Two adult cases of congenital atresia of the left coronary ostium-comparison of a sudden death case with a long-term survival case.

We present two extremely rare autopsy cases with atresia of the left coronary ostium. Case 1 was a 34-year-old man who died of sudden cardiac causes with localized acute subendocardial infarction and Case 2 was an 82-year-old woman who died from non-cardiac causes. In both cases, a string-like small left main trunk that was located in its normal position was found; however, the orifice of the left coronary artery was absent in both cases. The collateral artery (CA), whose caliber was smaller than that of the proximal left descending artery (LAD), arose from the right coronary sinus and was anastomosed with the distal LAD. However, it was only in Case 1 that the CA showed an acute angle of take-off from the aorta, subsequently coursing between the root of the aorta and the pulmonary trunk, with possible mechanical compression by these two arteries. These two cases suggest that atresia of the left coronary ostium is developmentally and anatomically different from single coronary artery or left coronary artery arising from the opposite sinus of valsalva. Atresia of the left coronary ostium is considered to be an important differential diagnosis as a cause of sudden death, not only in the pediatric population but also in adults. The caliber and/or other anatomical features of the CA may predominantly dictate the clinical course of patients with atresia of the left coronary ostium.

Polymorphisms of the cholecystokinin gene promoter region in suicide victims in Japan.

Cholecystokinin (CCK), a neurotransmitter in the central nervous system (CNS), co-exists in a large portion of A10 dopamine neurons to exert some effect on dopamine behavior. The aim of this study was to determine whether any association exists between the genotype of CCK gene promoter regions (-45C/T and -196G/A) and suicidal behavior. Genotypes and allele frequencies of CCK -45C/T and -196G/A were analyzed using polymerase chain reaction (PCR) followed by single-strand conformational polymorphism (SSCP) analysis on the genomic DNA from selected suicide victims (N=154) and from control subjects (N=328). Statistical analysis was performed using the Mantel-Haesnzel chi2-test and multiple logistic regression analysis with distinction of gender. An association between CCK -196G/A polymorphism and suicidal behavior in Japanese males was confirmed by statistical analysis (Odds ratio: 3.462, 95% CI: 1.128-10.626, P=0.038 by multiple logistic regression analysis). However, a significant association between CCK -196G/A polymorphism and suicidal behavior was not discovered in females. The polymorphism of the CCK gene promoter region was found to represent a susceptibility factor for suicidal behavior in Japanese males.

Risk factors of sudden death in the Japanese hot bath in the senior population.

A series of experiments were carried out to clarify the cause of death and the risk factors related to sudden death in the Japanese senior population while bathing in a Japanese style "hot bath." The biodynamic changes while bathing were carefully monitored under actual bathing situations occurring in both the winter and summer seasons. We observed double product (DP), total peripheral blood vessel resistance (TPR), cardiac output (CO), and blood vessel compliance (COMP) by measuring blood pressure, heart rate, pulse wave, and electrocardiogram (ECG). The finding of a high level of DP in the elderly suggests that more myocardial oxygen consumption is needed than for young adults, particularly in subjects with arrhythmia. Although the values for TPR and CO changed somewhat during bathing, the changes were considered normal and to be expected. However, more significant and substantial changes were observed during the winter experiment than during the summer experiment, no doubt owing to lower temperature of the bathing room. The value of COMP did not vary significantly between winter and summer subjects. Twelve subjects in the elderly developed ECG changes while bathing such as supraventricular extrasystole or ventricular tachycardia. No clinical significance was found in the biochemical analyses of the blood obtained before and after bathing. In conclusion, some subjects in the elderly showed risky changes in the above parameters and ECG, factors which may partially explain some of the causes of the many reported cases of lapse of consciousness and unexpected sudden death in the elderly while bathing especially in the winter season. Cold climate, hot water immersion, and hydrostatic pressure may affect their physiological compensation along with existing of coronary stenosis or weakness of respiratory function as a normal consequence of advanced age.

Sudden unexpected death of a 17-year-old male infected with the influenza virus.

We report a case of sudden unexpected death in a 17-year-old male student showing similar clinical background and pathological findings to Reye's syndrome. He was found following cardio-pulmonary arrest in his bed, and was immediately transferred to a hospital. However, resuscitation was not successful. He had a history of high fever of 38.3 degrees C, general malaise, myalgia, and gastrointestinal discomfort for the 2 days prior to his death, and an injection of pylazolon and medication comprising anti-emetics had been administered the day before he died. His biochemical findings showed almost normal levels of transaminase, electrolytes and protein fractions at the emergency room, but blood from the heart at autopsy revealed a high titer of the influenza A virus. Macroscopically, in addition to considerable fatty metamorphosis of the liver, concentric hypertrophy of the left ventricle, muscular bridge of left anterior descending artery, moderate coronary atherosclerosis, and mild downward displacement of the septal leaflet of the tricuspid valve were noted in the heart. Although panlobular microvesicular fatty infiltration of the liver was seen, deposition of lipid droplets was detected only in hepatocytes by frozen section of several organs. Serial sectioning of the epicardial coronary arteries showed about 50% stenosis at the distal site of the left circumflex artery, and diffuse interstitial fibrosis was evident in the bilateral ventricle and this was relatively severe for his age. In addition, the atrioventricular (AV) node artery showed severe narrowing just before entering the AV node, and downward displacement of the AV node with longitudinal elongation was also remarkable. We consider that the cause of death was sudden cardiac death rather than Reye's syndrome (RS), and that an arrhythmogenic event due to some preceding unusual cardiac lesions may have become overt due to the influenza infection and/or some related disorders. The present case would seem to suggest that a postmortem diagnosis of RS should be determined very carefully in cases of sudden death, even if the general circumstances would seem to be consistent with RS.

Comparison of age estimated from degree of racemization of aspartic acid, glutamic acid and alanine in the femur.

Aspartic acid (Asp) is generally used for estimation of age by measuring the degree of racemization. For other amino acids, however, there are few reports regarding the usefulness of the degree of racemization for the estimation of age. Accordingly, in this study using the femur (obtained from 21 cadavers) as the specimen, we measured the degree of racemization of glutamic acid (Glu) and alanine (Ala) along with Asp in the total amino acid (TAA) fraction as well as in acid-insoluble collagen-rich (IC) and acid-soluble peptide (SP) sub-fractions. We compared the degrees of racemization of each amino acid and the accuracy of the ages estimated from them. The degree of racemization and the reaction rate of racemization were ranked in the order of Asp > Glu > Ala in the TAA and IC fractions, but Asp > Ala > Glu in the SP fraction. It is noteworthy that the degrees of racemization differed between the three amino acids depending on the fraction tested. The correlation coefficient (r) between the degree of racemization and the chronological age was higher in the SP than in the TAA or IC fraction. Among three amino acids, Asp showed the highest correlation coefficient as predicted. The present study confirmed that Asp from the SP fraction is the best indicator for age estimation using racemization rates.

Radiographic survey of third molar development in relation to chronological age among Japanese juveniles.

The aim of the present study was to establish Japanese reference material on the third molar development of Japanese juveniles for forensic application. Observations were performed on the orthopantomograms of 1282 Japanese patients between the ages of 14.0 and 24.0 years. Demirjian formation stages of the maxillary and mandibular third molars were recorded for chronological evaluation of wisdom teeth and applied for further statistical analysis. Statistically significant differences were noted between the upper and lower jaws and genders. Accordingly, males achieved root developmental grades earlier than females. We assessed the mean ages for all formation grades and predicted the probability that a Japanese juvenile would be older than the relevant ages of 14, 16, and 20 as defined by Japanese Juvenile Law. We determined the likelihood that a Japanese youth is older than the relevant age of 18 as defined by legislation in the United States.

Age estimation from aspartic acid racemization of root dentin by internal standard method.

In this study, we investigated the application of the internal standard method to determine age from aspartic acid (Asp) racemization. D-Methionine (D-Met) and D-norleucine (D-Nleu) were tested as internal standards for the purpose of validating the derivatization and gas chromatographic measurements. Using a set of standard amino acids plus the internal standards in constant volume, calibration plots with reasonable linearity (R > 0.98) were constructed. Based on the analysis of sample chromatograms, D-Met appeared to meet the criteria for internal standards, hence it was selected for use in D- and L-Asp quantification. The correlation between dentin age and D-/L-Asp ratios from the peak areas as well as from the absolute concentrations was investigated. Correlation coefficients were calculated as 0.98 and 0.90, respectively. The slight decrease in accuracy was attributed to the conversion of D-Asp/D-Met ratios to concentrations employing the calibration curves figured from pure Asp. Because the application of the internal standard method produced reproducible and precise measurements, the employment of internal standards in age estimation based on Asp racemization appears to provide quality assurance by avoiding possible errors arising from sample preparation.

Polymorphisms of the tryptophan hydroxylase gene and serotonin 1A receptor gene in suicide victims among Japanese.

Akita Prefecture, Japan, has consistently recorded the highest level of suicide rates in all of Japan. In this study, we attempted to determine whether genetic differences between suicide victims and the normal population in Akita exist. We also researched the geographical differences in polymorphisms of the genes between people living in Akita Prefecture and those living in other prefectures with lower suicide rates as recorded in previously-published studies. Specifically, we investigated two serotonin-related genes including three substitutions connected to human emotional states such as despondency and depression: the tryptophan hydroxylase (TPH) gene (A779C and A218C in the intron) and the serotonin1A (5-HT1A) receptor gene (Pro 16Leu in the cording region). 134 suicide victims and 325 healthy volunteers were examined. For this process, we used two analytical procedures: (1) polymerase chain reaction (PCR) followed by single-strand conformational polymorphisms analysis for the A779C of TPH and the 5-HT1A receptor genes and (2) PCR followed by restriction fragment length polymorphism analysis for the A218C of TPH gene. No significant differences of the genotypes and the allele frequencies between the suicide samples and those of the healthy controls were discerned. Moreover, the genotype distributions of the TPH and 5-HT1A receptor genes were compared between Akita Prefecture and other prefectures, but no significant differences were found. In conclusion, no significant relation could be established statistically concerning the serotonin related genes between the suicide samples and control samples in Akita.

The examination of mushroom poisonings at Akita University.

In the past 10 years from 1991 to 2000, the number of consultations to the Japan Poison Information Center were 947 concerning mushroom poisonings. However, those from the hospital cases were not analyzed toxicologically. We examined toxicologically 20 cases (35 patients) of mushroom poisonings from 1993 to 2001. Investigation of amanita toxin poisoning was requested in 19 cases. We could detect the amanita toxin, amanitin, and phalloidin, in two cases, which resulted in concluding the cause of death. A fatal case by the magic mushroom poisoning was analyzed in the blood, urine, and mushroom, and we detected the hallucinogenic substances from the body fluids and ingested mushrooms. We report the results of our examinations, and point out the usefulness of the examination of the mushroom itself and biological samples toxicologically for forensic practice.

Forensic consideration of death in the bathtub.

About 100-120 cases of 'sudden death in bathroom' are reported every year in our prefecture, which is 8-10% of the total number of what is considered unnatural deaths. For reasons that are still unclear, the victims die while bathing in the bathtub. This baffling form of death occurs mostly in the winter season and 80-% of these are elderly persons. However, despite a full autopsy, it is sometimes still very difficult to determine the cause of death or the direct cause of drowning in the bathtub. To prevent others from this mysterious form of death and to determine the risk factors for the elderly while bathing, we examined several parameters such as physiological changes and biochemical blood analysis in a total of 54 volunteers during both the winter and summer seasons in Japan. We found that some of the elderly developed cardiac arrhythmia such as ventricular tachycardia, ventricular extrasystole, supraventricular extrasystole, and bradycardia. Also, decreases of systolic blood pressure during bathing were observed during this experiment. We conclude that the development of extrasysytole or ventricular tachycardia can be a stress involved in bathing. These changes might contribute to cardiac arrest or to drowning in the bathtub. Moreover, a sudden decrease of blood pressure is considered to be a risk factor of cerebrovascular accident.

Epitope analysis of human immunoglobulin D with a mouse monoclonal anti-Fab delta chain antibody.

BACKGROUND: The clinical significance of IgD measurements in serum is limited as the frequency of abnormal concentrations is rare. METHODS: We prepared a mouse monoclonal antibody and a rabbit polyclonal antibody against Fab delta and Fc delta chain and compared epitope recognition by the monoclonal antibody against Fab delta (anti-Fab delta mono) with that by other antibodies. RESULTS: Anti-Fab delta mono specifically reacted with purified IgD and Fab delta of myelomatous origin, but not with other isotypes and light chains. In 19 of 22 myeloma sera, the monoclonal antibody recognized intact IgD and/or its fragments when analyzed by immunoblotting. Of these there were only four cases in which possible Fab delta fragments were identified. The other three sera showed no reactivity with the antibody and the IgD value was low on a chemiluminescence enzyme immunoassay. CONCLUSIONS: These findings indicate the presence of at least two immunochemically different IgD molecules in the sera. No positive reaction with any synthetic peptide was observed for the antibody on delta-chain ranging from N-terminus of JH, C delta 1 to a hinge region. We suggest that the epitope recognized by the antibody is related to the variable region or a conformational structure on the Fd delta region of IgD.