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Interleukin (IL)-8 has been suggested to be a positive regulator of myelination in the central nervous system, in addition to its principal role as a chemokine for neutrophils. Immunostaining for beta-amyloid precursor protein (AβPP) is an effective tool for detecting traumatic axonal injury, although AβPP immunoreactivity can also indicate axonal injury due to hypoxic causes. In this study, we examined IL-8 and AβPP immunoreactivity in sections of corpus callosum obtained from deceased patients with blunt head injury and from equivalent control tissue. AβPP immunoreactivity was detected in injured axons, such as axonal bulbs and varicose axons, in 24 of 44 head injury cases. These AβPP immunoreactive cases had survived for more than 3h. The AβPP immunostaining pattern can be classified into two types: traumatic (Pattern 1) and non-traumatic (Pattern 2) axonal injuries, which we described previously [Hayashi et al. Int. J. Legal Med. 129 (2015) 1085-1090]. Three of 44 control cases also showed AβPP immunoreactive injured axons as Pattern 2. In contrast, IL-8 immunoreactivity was detected in 7 AβPP immunoreactive and in 2 non-AβPP immunoreactive head injury cases, but was not detected in any of the 44 control cases, including the 3 AβPP immunoreactive control cases. The IL-8 immunoreactive cases had survived from 3 to 24 days, whereas those cases who survived less than 3 days (n=29) and who survived 90 days (n=1) were not IL-8 immunoreactive. Moreover, IL-8 was detected as Pattern 1 axons only. In addition, double immunofluorescence analysis showed that IL-8 is expressed by oligodendrocytes surrounding injured axons. In conclusion, our results suggest that immunohistochemical detection of IL-8 may be useful as a complementary diagnostic marker of traumatic axonal injury.

In order to diagnose death associated with fire, it is essential to show that the person was exposed to heat while still alive. We investigated both AQP1 and AQP3 expression in the skin of an experimental burn model, as well as in forensic autopsy cases, and discuss its role in the differential diagnosis of ante- and postmortem burns. In animal experiments, there was no difference in AQP1 gene expression among four groups (n=4): antemortem burn, postmortem burn, mechanical wound, and control. However, AQP3 expression in the antemortem burn was increased significantly compared with that of the other groups even at 5min after burn. Water content of the skin was decreased significantly by the burn procedure. Consistent with animal experiments, AQP3 gene expression in the skin of antemortem burn cases was increased significantly compared with postmortem burns, mechanical wounds, and controls (n=12 in each group). These observations suggest that dermal AQP3 gene expression was increased to maintain water homeostasis in response to dehydration from burn. Finally, our results suggest that AQP3 gene expression may be useful for forensic molecular diagnosis of antemortem burn.

Determination of the age of burns, as well as of wounds induced mechanically, is essential in forensic practice, particularly in cases of suspected child abuse. Here, we investigated temporal changes in the expression of 13 genes during wound healing after a burn. The expression of cytokines (IL-1β, IL-6, IL-10, TNF-α, and IFN-γ), chemokines (KC, MCP-1), proliferative factors (TGF-β, VEGF), proteases (MMP-2, 9, 13) and type I collagen in murine skin was examined by real-time PCR at 3, 6, 9, and 12 h and 1, 2, 3, 5, 7, and 14 days after a burn. Based on macroscopic and histological appearance, the healing process of a burn consists of 3 phases: inflammatory (from 3 h to 1 day after the burn), proliferative (from 1 to 7 days), and maturation (from 7 to 14 days). Expression of IL-1β, IL-6, TNF-α, IFN-γ and KC increased significantly in a biphasic pattern from 3 or 6 h to 12 h or 1 day and from 3 or 5 days to 7 days. Expression of MCP-1 increased significantly from 6 h to 5 days. Expression of both IL-10 and TGF-β increased significantly from 12 h to 7 days. Expression of VEGF, MMP-2, MMP-13 and type I collagen increased significantly from 3 days to 7 or 14 days. Expression of MMP-9 increased significantly from 6 h to 14 days. Our results suggest that evaluating the expression of a combination of these genes would enable the exact estimation of the age of a burn.

We report a forced double suicide involving a wife and husband in their late 80s resulting from chloroform inhalation. Macro- and microscopic examinations revealed marked pulmonary edema and extensive contraction band necrosis of the cardiac muscles. Toxicological analysis revealed high levels of chloroform in the blood (41.4μg/ml in the wife and 29.1μg/ml in the husband) and in the adipose tissue (128μg/g in the wife and 131μg/g in the husband). From these findings, we conclude that the cause of death of both was acute heart failure due to chloroform poisoning. In addition, the pathological examination of the husband revealed submucosal hemorrhage at the root of the tongue and trachea, erosion of the stomach, and upper jejunum, none of which were present in the wife. Furthermore, hyperemia of the mucous membrane of the husband was more marked than that of the wife. Toxicological analysis also revealed that the chloroform levels in the liver and brain of the husband were higher than those of the wife, although the chloroform level in the blood of the husband was lower than that of the wife. We presume that the wife may have inhaled a greater amount of chloroform, and that the wife's circulation may have arrested before irritation of the mucous membranes became apparent. The husband may have inhaled a smaller amount of chloroform in longer duration, leading to irritation of the mucous membranes prior to the fatal heart failure. These findings suggest that toxicological and pathological outcomes of chloroform poisoning may vary between patients, and that they may reflect the dose and duration of chloroform inhalation.

The infiltration of 'primed' polymorphonuclear neutrophils into multiple organs has been reported in cases of traumatic or hemorrhagic shock. Since multiple injuries are usually observed in cases of physical abuse of the elderly, we investigated neutrophil infiltration into the heart, lung, liver and kidney in cases of abused elderly individuals using immunohistochemistry for myeloperoxidase (MPO). In addition, we examined the expression of molecules associated with neutrophil infiltration, including P-selectin as the adhesion molecule and IL-8 as the chemotactic factor. The number of neutrophils in the physically abused elder cases was increased significantly, particularly in the lung and liver, compared with that of control cases of sharp instrument injury, single fatal blunt injury and polytrauma. In addition, P-selection expression in the endothelium and the presence of IL-8-positive cells (mainly macrophages) in the lung and liver of abuse cases were significantly greater than those in control cases. In contrast, the number of MPO-, P-selectin- and IL-8-positive cells in cases of multiple organ failure (MOF) due to various causes was significantly greater than that in abuse cases. It is known that primed neutrophils accumulation may undergo MOF by 'activation' due to secondary insults. Thus, our results suggest that MPO immunostaining can distinguish cases of elderly physical abuse from non-abuse and MOF cases. In addition, our results indicate that MPO is a potential diagnostic marker for elder physical abuse, and that P-selectin and IL-8 may be useful for a more accurate diagnosis. Finally, our results also suggest that elder cases of physical abuse may be in a primed stage of MOF, and are at risk of falling into MOF by various secondary insults including those following abuse.

We describe an unusual case of total replacement of the exocrine pancreas with fat, which was observed in an autopsy of an assaulted victim. A woman in her early 80s was kicked, stamped and hit several times with firewood. She was hospitalized with disturbance of consciousness, left haemothorax and multiple fractures, and died about three months later. Postmortem examination revealed extensive abrasions and bruises, multiple fractures and internal organ injuries such as contusion and haemorrhage, as well as bronchopneumonia. It was concluded that the cause of her death was hypostatic pneumonia followed by traumatic shock due to multiple blunt injuries. Further, complete replacement of the pancreas with fat was observed in addition to a calculus in the main pancreatic duct and fibrous hypertrophy of the ductal wall. Histopathological examination revealed almost complete replacement of the pancreatic acini by fat tissue, whereas the islets of Langerhans were mostly intact. Antemortem laboratory data showed that serum amylase levels were almost within normal range before hospital admission, but underwent a transient abnormal elevation at admission followed by extremely low levels thereafter. Previous reports suggest that obstruction of both the main pancreatic duct and the artery, due to tumour formation or calculus in combination with arteriolar sclerosis, are necessary to induce total replacement of the pancreas with fat. Since arteriolar sclerosis was not remarkable in this case, we speculated that pancreatic ischaemia due to circulatory disturbance caused by traumatic shock, in combination with pre-existing calculus, may have contributed to the development of total replacement with fat. The temporal alterations in serum amylase levels support our speculation. There are few, if any, reports regarding organ replacement with fat in association with trauma. This case suggests that multiple injuries followed by traumatic shock may advance pre-existing replacement of the pancreas with fat.

Immunostaining for beta-amyloid precursor protein (APP) is recognized as an effective tool for detecting traumatic axonal injury, but it also detects axonal injury due to ischemic or other metabolic causes. Previously, we reported two different patterns of APP staining: labeled axons oriented along with white matter bundles (pattern 1) and labeled axons scattered irregularly (pattern 2) (Hayashi et al. (Leg Med (Tokyo) 11:S171-173, 2009). In this study, we investigated whether these two patterns are consistent with patterns of trauma and hypoxic brain damage, respectively. Sections of the corpus callosum from 44 cases of blunt head injury and equivalent control tissue were immunostained for APP. APP was detected in injured axons such as axonal bulbs and varicose axons in 24 of the 44 cases of head injuries that also survived for three or more hours after injury. In 21 of the 24 APP-positive cases, pattern 1 alone was observed in 14 cases, pattern 2 alone was not observed in any cases, and both patterns 1 and 2 were detected in 7 cases. APP-labeled injured axons were detected in 3 of the 44 control cases, all of which were pattern 2. These results suggest that pattern 1 indicates traumatic axonal injury, while pattern 2 results from hypoxic insult. These patterns may be useful to differentiate between traumatic and nontraumatic axonal injuries.

The species specificity of 10 human Y chromosomal short tandem repeat loci, DYS19, DYS385, DYS389, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439, was examined in the Japanese macaque, Macaca fuscata. Primers for loci DYS391 and DYS438 only yielded an amplification product from male samples. The PCR products amplified with the DYS391 primers showed no inter-individual differences in migration rate in electrophoretic experiments. Sequence analysis revealed that these PCR products consisted of 287 bases containing tandem repeats of TC and TATC. The numbers of TC and the TATC repeats varied between individuals. The TC repeat does not exist in human and chimpanzee sequences. The PCR products amplified by the DYS438 primers provided no evidence of inter-individual variation between the six male Japanese macaque samples. In the Japanese macaque, PCR gives a 184 base pair product, in contrast to human sample from which the products are 203-233 bases in size. The primers for four loci, DYS19, DYS385, DYS389 and DYS437 produced PCR products from both male and female Japanese macaques. The primers for the other loci, DYS390, DYS392, DYS393 and DYS439, did not yield PCR products.

We report a case of sudden death due to n-butane poisoning after the inhalation of anti-perspiration aerosol deodorant. The deceased was a 15-year-old boy who was found unresponsive on the road, and was pronounced dead after 1.25h. A spray can of anti-perspiration deodorant and vinyl bags were found in a thicket near the scene. An autopsy revealed pulmonary edema, cerebral edema and congestion of the organs. Using qualitative gas chromatography/mass spectrometry, the existence of n-butane was ascertained. The concentration of n-butane (in microl/ml or microl/g) was estimated to be 15.3 in the blood, 13.3 in the brain, 26.6 in the liver, 7.5 in the lung, and 13.6 in the kidney. These n-butane levels in the blood and in the tissues were higher than those of previous reports of death associated with n-butane inhalation. We concluded that the cause of death was n-butane poisoning and presumed that n-butane in the can of anti-perspiration aerosol deodorant induced fatal cardiac arrhythmia.

In this study alpha-1-antitrypsin (AAT) phenotypes at the protease inhibitor (PI) locus were determined by isoelectric focusing of native and desialylated serum samples from 236 Japanese subjects living in the western part of Japan. The shifts in relative mobility between some PI types were observed before and after desialylation. This technique was useful in distinguishing between some PI M subtypes and variants. The molecular basis of four variant alleles, including two new alleles found in this study, was characterized: PI E tokyo [Lys 335 ( A AG)→ Glu( G AG)] and PI N nagato [Leu 276 (C T G)→Pro(C C G)] arose from PI M1(Val 213 ) and PI M2, respectively. A new PI P yonago [Asp 19 (G A T)→Ala(G C T)] originated from PI M1(Val 213 ). A new PI M5 gunma [Pro 362 ( C CC)→Ser( T CC)], arising from PI M3, was the sixth allele involving a mutation at codon 362, which is suggested to be a mutation hot spot. PI M5 gunma was likely to show normal AAT levels and function although the mutations occurred near codon 358 for Met 358 . The molecular basis of PI variant alleles found in Japanese was different from that reported in previous studies.

As part of our comprehensive genetic study of the aboriginal Taiwan ese(Gaoshan), we have reported the distribution of allele frequencies for 13 red cell enzyme and 4 serum protein polymorphisms in all nine aboriginal groups. In this study, 10 additional serum protein polymorphisms for AHSG, C3, C6, C7, GC, HP, ITIH1, PI, TF and ZAG were typed. Common and variant alleles prevail ingin East Asian populations were found together with several new variants. The distribution of allele frequencies differed markedly among the nine aboriginal groups and from that in East Asian populations. The large diversity among the aboriginal groups was confirmed by analysis of the genetic affinity on the basis of Nei's standard genetic distance and Cavalli-Sforza's chord distance, which were calculated using the present and previous data from 27 red cell enzyme and serum protein loci. The analysis based on 15 red cell enzyme and serum protein loci revealed that they, in particular the Puyuma and Ami, had strong affinity with Filipino among several East Asian populations.

Immunostaining for beta-amyloid precursor protein (APP) is widely recognized as an effective tool for detecting diffuse traumatic axonal injury (TAI). APP selectively labels injured axons, such as axonal bulbs and varicose axons. However, it has been reported that axonal bulbs are detected in cases of cerebral hypoxia without head injury. Therefore, we examined whether there are differences in the morphological pattern of axonal bulbs between trauma and hypoxia. Sections of the corpus callosum from 25 cases of head injury and 23 control cases were immunostained for APP. APP staining detected axonal bulbs in 14 cases of head injury, who survived more than several hours, although it failed to label axons in control cases. In addition, two patterns of immunoreactivity were identified in several cases of head injury. The first pattern showed that labeled axons were oriented along with white matter bundles; the second demonstrated that the axons were scattered irregularly. The first pattern alone was found in 5 of 14 cases, while cases of the second pattern alone were not observed. Both patterns were detected in 5 cases and in the remaining 4 cases, clear patterns were not found. From these findings, we speculated that the first pattern may represent TAI. Further examinations are required for determining whether these two patterns are identical with patterns of trauma and hypoxic brain damage as indicated by [Oehmichen M, Meissner C, Schmidt V, Pedal I, König HG, Saternus KS. Axonal injury--a diagnostic tool in forensic neuropathology? A review. Forensic Sci Int 1998;95:67-83] and [Graham DI, Smith C, Reichard R, Leclercq PD, Gentleman SM. Trials and tribulations of using beta-amyloid precursor protein immunohistochemistry to evaluate traumatic brain injury in adults. Forensic Sci Int 2004;146:89-96].

To scrutinize the geographical clines of allele frequencies of the group-specific component/vitamin-D binding protein (GC) system in Japan, the GC types in nine populations of southwestern Japan including Okinawa were determined. The frequencies of the common alleles in the island populations, especially, Yakushima and Miyako were found to be scattered more extensively than those in the populations of the mainland Japan. GC*1A2 was polymorphic in all populations except in the southwesternmost islands, Miyako and Ishigaki. The geographical clines of GC*1F, GC*1S and GC*2 frequencies are discussed in connection with the migration of the Yayoi era.

The distribution of transferrin phenotypes was investigated in nine populations of southwestern Japan: mainland Kagoshima Prefecture and the Nansei Islands. Populations from Tanegashima to Okinawa were homogenous in the distribution of TF C subtypes. Miyako and Ishigaki in the southwesternmost region had higher frequencies of TF*C1, and lower frequencies of TF*C2 than the seven other populations. A new TF D variant, tentatively designated TF Dama, differed in isoelectric point from adjacent variants including TF Dchi, TF Doki and TF D1. TF*Dama was found to be a new genetic marker for the Nansei Islands. Geographical clines of TF allele frequencies are also discussed.

We examined the species specificity of six commercially available human short tandem repeat systems (for CSF1PO, TPOX, TH01, F13A01, FESFPS and vWA) using six samples obtained from the Japanese monkey Macaca fuscata. Macaque genes were amplified with human TPOX and F13A01 primers. During electrophoresis, macaque polymerase chain reaction (PCR) products amplified with the TPOX primer migrated more slowly than human ones, and migrated close to human CSF1PO bands. Macaque PCR products amplified with the F13A01 primer also migrated more slowly than the human equivalent. The six macaque samples were classified into three types by the TPOX primer and into six types by the F13A01 primer. These results suggest that the primers for human TPOX and F13A01 loci can be used to distinguish between human and Japanese macaque samples.

Endogenous glucocorticoid-induced thymic involution is generally considered to be an important finding for determining child abuse. The present study investigated the weight of the thymus and the adrenal glands in elder abuse cases to identify a potential marker for elder abuse. There was no significant difference in the thymus and the adrenal weight between elder abuse and control cases. However, the elder abuse cases in which the duration of abuse was less than 3 months showed a significant increase in the adrenal weight in comparison to control cases. In such cases, histopathological findings showed a loss of intracellular light granules from the zona fasciculata, which might indicate a loss of cholesterol due to the overproduction of glucocorticoid. These results might imply that the elderly, who were maltreated for less than 3 months, were in the early phase of a long-term stress state during which stress-induced overproduction of glucocorticoid was observed in adrenal glands as indicated by Selye. Our results suggest that an increase in adrenal weight may be a potential marker for elder abuse of relatively short periods, especially less than a few months.

The case of a monozygotic twin, discordant for congenital hypothyroidism, is reported. The monozygosity of the twins was established by typing for 23 different hematogenetic marker gene-systems, with DNA fingerprint analysis. The sublingual thyroid in this patient was probably due to an intrauterine disorder of unknown origin which spared his brother.This case supports the hypothesis that environmental, rather than genetic factors, are the cause of ectopic thyroid dysgenesis.

We describe an autopsy case in which a patient with diabetic ketoacidosis (DKA) was found in a head-down position. A female in her late 70s was found dead in her home in a supine position on the kitchen floor. The upper part of her body was hanging down over the edge of the kitchen floor to the backyard through the open window. External examination revealed congestion of the head and upper region of the face and neck. There were numerous petechiae on the superior palpebral conjunctivae and upper part of the oral mucosa. On internal examination, extensive hemorrhages in the subcutaneous fat tissues and muscles were observed at the upper part of the neck, although there were no external injuries on the neck. Histopathological examination revealed that hemorrhages were accompanied with infiltration of polymorphonuclear leukocytes both within and around the hemorrhages on the neck skin. Nodular glomerulosclerosis and many fat droplets in the cytoplasm of proximal tubule cells were found in the kidney. Postmortem blood analysis showed acetone (204.2 μg/ml), HbA1c (10.8%), acetoacetate (

In Japan, sudden death in the bathroom (what we call ‘bath-related death’) has been reported to occur particularly in the elderly population in the winter. This investigation aimed to obtain a better understanding of the factors associated with bath-related deaths. For this purpose, a retrospective review of the inquest records in Kagoshima Prefecture, the southwest part of Japan, during 2006–2007 was performed. There were 338 cases of bath-related death, which corresponds to a crude mortality rate of 9.7 per 100,000 persons per year. The occurrence of bath-related death was similar to that of other prefectures in Japan, despite the warm environment in Kagoshima. The victims' ages ranged from 37 to 101 years (mean 76.6 years) and 88.8% of patients were over 65 years. The associated factors included winter season (56.2%) and a medical history of hypertension (37.3%). These demographic and circumstantial factors correlated with previous studies in Japan. These results suggest that the Japanese method of bathing, in addition to cold bathrooms in winter, contributes to the high rate of bath-related death in Japan. In addition, the mortality rate of people who lived alone was significantly higher than that of those who lived with their families. Most cases occurred in the home bathroom (84.9%) at the time when elderly people usually bathe (48.7%). Our results indicate that bath-related death occurs most often during the normal daily life of the aged. Protective activities of the Japanese government and society should aim to reduce the number of these tragic deaths in the bathroom. Further, no autopsies were performed on the cases in this study. Therefore, the rate of autopsies in bath-related death should be increased in order to accumulate data based on objective evidence.

We assessed the plankton levels using the diatom test in the lungs and organs of the circulatory system of nine postmortem cases of bathwater drowning and one case of death due to ischemic heart disease (IHD) while bathing, as well as in the associated bathwater. The number of planktons detected in the lungs was not related to the postmortem period or the process of drowning, but was related to the plankton levels detected in the bathwater taken from the scene. In one case of bathwater drowning, the diatom distribution of the four pulmonary lobes (165.0–280.0planktons/100g sample) and other organs (3.6–13.6/100g) highly corresponded to the pattern of ‘usual' drowning in water which contains a lot of diatoms. In this case, the bathwater taken from the scene contained a considerable amount of planktons (16.3/100ml water), which suggests that, when the bathwater is relatively ‘dirty', the diatom test may be helpful in reaching a diagnosis of drowning. In another case of drowning, a considerable number of diatoms was detected relatively evenly in all four pulmonary lobes (58.3–141.7/100g), although no planktons were detected in distant organs. In this case, the bathwater also contained a substantial number of planktons (18.8/100ml), which suggests that a relatively even distribution of planktons in each pulmonary lobe may also support a diagnosis of drowning based on the autopsy findings when the bathwater is relatively ‘dirty'. In the other seven cases of drowning, few planktons were detected in the lungs (0–18.2/100g) and other organs (0–9.1/100g) as well as in the associated bathwater (0–3.1/100ml). These cases suggest that if autopsy findings indicate drowning, the low levels of diatoms detected in both the pulmonary lobes and the bathwater may indicate drowning in clean water. In contrast, in one case of death due to IHD while bathing, no planktons were detected in the organs except for the lower lobe of the right lung (11.8/100g) and the left kidney (9.1/100g), although the bathwater contained a sufficient number of planktons by the diatom test (21.3/100ml). This case suggests that the diatom test may be helpful in distinguishing drowning and other causes of death in bathwater by comparison of the diatom levels in the organs and the bathwater. All of these results indicate that the diatom test may be helpful to differentiate ambiguous cases of drowning in bathwater. However, in this study, although special care and precautions were undertaken, contamination with some planktons could not be prevented. For accurate interpretation and diagnosis, examination and comparison of test results from all four pulmonary lobes, at least four distant organs, and the associated bathwater from the scene of death are necessary together with careful consideration of autopsy findings.

A male in his early seventies complained of abdominal pain and diarrhea at 7h after ingesting a small piece of gratin from a box lunch prepared by a caterer. He was admitted to a hospital, but died 37 h later. Dozens of people who had eaten the same box lunch also complained of diarrhea. All of them recovered after medical treatment. A later investigation demonstrated Salmonella Enteritidis (SE) in the gratin from the box lunch. An autopsy revealed very severe typhloenteritis with edema and submucosal hemorrhage. The digestive tract contained fluid contents without foodstuffs. Bacteriological examination revealed SE in the contents of the lower ileum and large intestine. Based on these findings, we concluded that the cause of death was food poisoning due to SE. In this case, ingesting only a small piece of contaminated food caused fatal food poisoning due to SE. These results emphasize the importance of prevention against food poisoning due to Salmonella, particularly SE.

Penta E and D18S51 loci are known to be highly polymorphic in humans. In this study, Japanese macaque PCR products amplified with human Penta E and D18S51 primers were examined. Electrophoresis and sequence analyses revealed that 13 Japanese macaque products amplified with human Penta E primer were homogeneous. The Japanese macaque sequence was extremely similar with human Penta E alleles except for the insertion of contiguous adenosine repeats, called ‘A-stretch', at 5′-side of AAAGA repeats. Among the 11 Japanese macaque PCR products amplified with human D18S51 primers, only two variants were observed. The sequences of these Japanese macaque products were similar to those of human D18S51 alleles. However, the Japanese macaque sequences also contained the insertion of A-stretch at 5′-side of AGAA repeats. Less polymorphism in the Japanese macaque sequences, in contrast with highly polymorphic human Penta E and D18S51 loci, suggested that A-stretches might inhibit the emergence of new variants at the STR loci.

In a morning in January, a male in his early sixties was found dead in an outdoor parking area. The minimum temperature during the night before he was found dead was estimated to be 4.0 degrees C. Autopsy revealed the pinkness of hypostasis, slight abrasions and bruises on the face and the extremities, collapse of the lungs, and slight gastric submucosal hemorrhage. Histologic examination revealed compact arrangement of cardiac muscle fibers and cytoplasmic vacuolation in the adenohypophysis. Toxicologic examination demonstrated hyperacetonemia (51.2 microg/mL). Ubiquitin, one of the stress proteins that are induced by several stimuli, including severe cold, was detected in several organs. We concluded that the cause of his death was lethal hypothermia. In addition, hemorrhages were observed in the subfascial and/or intramuscular parts of the pectoralis minor, first intercostal, and iliopsoas muscles. Although it has been reported that iliopsoas muscle hemorrhage can result from hypothermia, there have been few reports concerning hypothermia-associated hemorrhages of the pectoralis minor and/or intercostal muscles. We presumed that intense shivering and/or effort ventilation during the course of lethal hypothermia might cause these muscle hemorrhages.

We report the simultaneous deaths of two individuals by inhalation of hydrogen sulfide (H(2)S), produced either by the putrefaction of a large quantity of sweet corn or by heavy oil that flowed out of the fuel tank of a large stranded cargo vessel. Ten workers went into a water ballast tank of the vessel to remove remaining heavy oil and suddenly felt unwell. Two of the ten workers (patient A, a male in his early thirties, and patient B, a male in his early sixties) died. Autopsies of the two patients revealed a partial green discoloration of the skin and pulmonary edema. Toxicological analysis revealed high levels of thiosulfate in the blood (0.089 mmol/L in patient A and 0.142 mmol/L in patient B). From these findings, we concluded that the cause of death in both patients was H(2)S poisoning. In addition, the autopsy of patient A revealed petechiae of the palpebral conjunctiva and the mucous membrane of the mouth and erosion of the respiratory tract. The autopsy of patient B failed to reveal these observations. We presumed that patient B may have been exposed to higher H(2)S levels, and that the circulation and respiration of patient B may have arrested faster than patient A. Thiosulfate levels in the blood may reflect the levels of H(2)S exposure. This case suggests that the pathological and toxicological findings of H(2)S poisoning vary from case to case.

Amplification products from male and female Japanese macaques were obtained by PCR with human Y-chromosomal DYS389 primers. These products were examined by electrophoresis and sequence analysis. The PCR products from the 12 Japanese macaques tested had different band patterns on an electrophoretogram. Sequence analysis of the products revealed that the high polymorphism originated from variable numbers of repeats of two separate CTAT sequences. The sequences of the Japanese macaque products were similar to those of the reference human DYS389 sequence. However, variable CTGT repeats and a difference in the second forward primer binding site yielded two products in human males, DYS389I and DYS389II, which do not exist in Japanese macaques. Our results suggest that the human DYS389 primers may be a potential tool not only for distinguishing between human and Japanese macaque DNA samples, but also for identifying individual macaques, because of the highly polymorphic alleles.

A man in his late twenties collapsed shortly after intravenously injecting himself with methamphetamine (MA). He slipped into a deep coma and remained in this condition for 9 days, until his death. Autopsy revealed severe brain edema and localized subarachnoid hemorrhages in the cerebrum and cerebellum. Histopathological examination revealed myocardial necrosis in the left ventricle, rhabdomyolysis and bronchopneumonia. Blood derived from the cadaver was found to have high levels of blood urea nitrogen and creatinine, suggesting he experienced acute renal failure probably due to rhabdomyolysis. Most of the postmortem findings were consistent with MA poisoning. The patient's bronchopneumonia may have represented a hypostatic pneumonia that developed as a result of his deep coma. While the patient's brain edema, myocardial necrosis and rhabdomyolysis were diagnosed soon after admission, his bronchopneumonia and acute renal failure only occurred 6 and 8 days later, respectively. Although MA was not detected in the cadaver's blood, urine or liver, analysis of the decedent's hair using gas chromatography-mass spectrometry confirmed its presence at a concentration of 1.1 ng/mg. Based on these findings, we concluded that the patient's cause of death was multiorganopathy resulting from MA poisoning. This case suggests that the postmortem diagnosis of MA poisoning in patients who survive for relatively longer periods after drug injection should include toxicological hair analysis in combination with histopathological and postmortem physiochemical examination.

We report an autopsy case of a death associated with ingestion of liquid windshield-washer detergent. The deceased was a 49-year-old man who was found dead on a road near his truck. A bottle of liquid windshield-washer detergent containing an anionic surfactant and methanol was found under the passenger seat of the truck. At autopsy, slight abrasions and bruises were observed on his body. The small intestine contained dark greenish-brown mucoid matter with abundant froth. The mucous membranes of the esophagus, stomach and superior small intestine showed extensive necrosis, erosion, hemorrhage, edema and congestion. Using the methylene blue method to examine the contents of the small intestine, the presence of an anionic surfactant was indicated. We conclude that the cause of death was ingestion of liquid windshield-washer detergent.

We examined six human short tandem repeat (STR) loci (c-fms proto-oncogene for CSF-1 receptor, CSF1PO; thyroid peroxidase, TPOX; tyrosine hydroxylase, TH01; coagulation factor XIII a subunit, F13A01; c-fes/fps proto-oncogene, FESFPS and von Willebrand factor, vWA) in the Japanese macaque, Macaca fuscata, using commercially available human STR kits. No products were amplified by polymerase chain reaction (PCR) using the human CSF1PO, TH01, FESFPS and vWA primers. Macaque DNAs were amplified with human TPOX and F13A01 primers. Macaque PCR products amplified with the TPOX and F13A01 primers migrated more slowly than human ones during electrophoresis. Sequencing results showed that the nucleotide sequences of the counterpart of the TPOX and F13A01 STR loci in the Japanese macaque were closely similar to those of the human genes except for tandem repeat regions. The macaque products amplified with human TPOX and F13A01 primers were highly polymorphic, with four variants of the former and 15 variants of the latter in the nine samples. These results indicate that the commercially available kits can be used to discriminate the Japanese macaque samples from human samples.

A number of mutations in coding and noncoding regions of mitochondrial DNA (mtDNA) have previously been studied. In the present study, we simultaneously typed six mutation sites in the coding region by use of amplified product-length polymorphism (APLP) analysis. The mtDNA variations of 2471 individuals from 20 populations of Japanese, Korean, Chinese, and German were examined and classified into 18 haplotypes. Two of these haplotypes, B1 (estimated ancestral haplotype) and C1, were distributed among all populations tested. However, the haplotypes A1, A2, B2, B3, and C2 were mostly restricted to the Mongoloid populations, whereas haplotypes B5 and C5 appeared almost exclusively in the German population. Phylogenetic analysis by the neighbor-joining method revealed that the Japanese populations were more closely related to each other than to the other East Asian populations surveyed. The multiplex APLP method is suitable for large-scale screening studies of mtDNA variability because it is both rapid and economical.

A female in her late thirties, who was hospitalized for 13 days with rib fractures suffered in a traffic accident, was found in supine position on the patient's bed in a state of cardiopulmonary arrest. The resuscitation failed to recover consciousness and her death was confirmed 1.75 h later. The autopsy revealed neither severe injuries nor lesion as a cause of death. Histological examinations revealed contraction band necrosis (CBN) in the cardiac muscle of the left and right ventricles, as well as congestion and edema in both lungs. Toxicological screening of urine using Triage revealed the presence of methamphetamine (MA). By gas chromatography-mass spectrometry (GC/MS), the concentrations of MA and its metabolite amphetamine (AP) in the heart blood were shown to be 0.178 and 0.051 microg/ml, respectively. From these findings, we conclude that the cause of her death was circulatory failure due to MA poisoning. Further, the stored scalp hair analysis demonstrated both MA and AP in 0-3 cm segments from the scalp. The results suggested that she was a MA abuser at least about 3 months before her death. In this case, preservation of hair samples collected during an autopsy was useful for providing evidence of MA abuse.

ブタの白血球型分類および抗血清の標準化のため,著者およびフランス人のVAYMANらが豚同種皮膚移植免疫によって分類作製した合計23種類(59例)の抗リンパ球型血清について比較同定試験を行った.抗豚リンパ球型血清はいずれも細胞毒抗体(cytotoxic antibody)であり,その抗体価は16倍-8,192倍であった.また,25例の豚リンパ球panel cellsを用いて,リンパ球細胞毒試験(lymphocytotoxity test)によってリンパ球型反応パターンを得たのち,抗血清相互の同定試験を行ったところ,N1•N2•N3•N10系の抗血清は抗SL-A4を,またN6•N7系の抗血清は抗SL-A1をそれぞれ一部共通特異性抗体として含有し,さらにN4は抗SL-A2,N8はSL-A未分類抗体(No. 1526)とほぼ同一の特異性を示す抗血清であった.しかし,N5およびN9はSL-A系抗体の17種類には適合せず,新たに分類された抗豚リンパ球型抗体と考えられる.