Mihoko Ago

Temporal expression of wound healing-related genes in skin burn injury

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.

A fatal case of n-butane poisoning after inhaling anti-perspiration aerosol deodorant

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.

Two fatalities by hydrogen sulfide poisoning: Variation of pathological and toxicological findings

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.

Two patterns of β-amyloid precursor protein (APP) immunoreactivity in cases of blunt head injury

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].

Positional asphyxia or diabetic ketoacidosis? A case report

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 (<2.0 μmol/l), 3-hydroxybutyrate (11,844 μmol/l), blood urea nitrogen (128.9 mg/dl), and creatinine (3.11 mg/dl). The glucose and acetone levels in the urine were 876.7 mg/dl and 201.4 μg/ml, respectively, suggesting that she suffered severe DKA. However, since hemorrhage of the neck could have developed only when she was still alive, asphyxia should have arisen antemortem. Based on these findings, we concluded that the direct cause of her death is positional asphyxia, which was resulted from DKA. It is difficult to diagnose the cause of death when the victim is in an unusual posture. To confirm a suspicion of positional asphyxia, photographs of the undisturbed scene are useful in addition to a precise autopsy and accurate examinations.

Forensic diagnosis of ante- and postmortem burn based on aquaporin-3 gene expression in the skin

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.

Thymus and Adrenal Glands in Elder Abuse

Abstract 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.

A case of death associated with ingestion of liquid windshield-washer detergent

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.

Determination of methamphetamine in sudden death of a traffic accident inpatient by blood and hair analyses.

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 patients 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.

Y-chromosomal STR haplotyping in a Japanese population

The Y-specific STR systems including DYS19, DYS385, DYS389I, DYS390, DYS391 and DYS393, were investigated in 117 Japanese males. Analysis of the 6 loci permitted classification of the samples into 90 haplotypes, and the haplotypic diversity was estimated to be 0.984. Distribution of the haplotypes in the Japanese population studied was different from that in European populations.