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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.

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.

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].

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.

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 (

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.

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.

Diffuse types of traumatic brain injury (TBI) are more difficult to diagnose than focal types in forensic postmortem examination, since macroscopic abnormalities may be minimal. In addition, most microscopic findings are not specific to TBI and are sometimes not obvious in cases when the survival period is short. Therefore, early diagnosis of diffuse TBI is most difficult. Histopathological and immunohistochemical examinations of various elements including axons, nerve cells, and glial cells in a sufficient number of blocks are indispensable. Mapping of changes in these elements with complicated focal lesions, even if the lesions are trivial, on anatomical diagrams would be useful. The combination of histopathological and immunohistochemical examinations as well as analysis of the exact history of the trauma, if possible, and elimination of other causes of death would lead to accurate diagnosis of diffuse types of TBI in cases when the survival period is brief.

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.

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.