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This paper presents an unusual complex suicide case that died of nicotine addiction. The deceased was a 40-year-old male who was found lying dead on the floor in his room. In external findings, many incision wounds on his forearms and skin discoloration with epidermolysis on his cervical region could be seen. In the room, a blood-stained scissors and electric cord hanged on the exercise bike were found. Moreover, nine cigarette residues which were only the filter part and empty bottle of coffee were found on his side. At autopsy, we found that those injuries were not serious enough to lead him to the death. Toxicologically, caffeine, nicotine, cotinine, mirtazapine, and olanzapine could be detected, and the concentrations of nicotine were 3.740, 2.140, 3.100, and 451.100 μg/ml in cardiac blood, peripheral blood, urine, and stomach contents, respectively. These concentrations were evaluated as the fatal levels, and the cause of his death was diagnosed as acute nicotine intoxication.

We report a suicide case of complete decapitation using a self-constructed guillotine. A 45-year-old man, whose body was severely burned, was found dead. The head was completely separated from the middle level of the neck, and a sharp blade with a steel frame was placed between the head and neck. The severance plane passed between the C4 and C5 vertebrae. Vital reactions such as hemorrhage could not be confirmed at the decapitated skin edge because the body was severely burned. Both common carotid arteries were sharply transected. Subendocardial hemorrhage was detected in the left ventricle. Only a little blood, but no soot, was detected in the respiratory tract, including the trachea and bilateral bronchi. Subarachnoid hemorrhage was noted at the edge of the cervical spinal cord. The saturation level of CO-Hb was 5.7% in the left cardiac blood, 5.9% in the right cardiac blood, and 5.8% in the peripheral blood from the femoral vein. Cervical transection was diagnosed as the cause of death. We believe that he was unintentionally burned by spread fire from an automobile after decapitation by a self-constructed guillotine.

•This report an intoxication case caused by the ingestion of the pesticide Ortoran®, which consists of 50% acephate.•Based on these results and autopsy findings, the cause of death was acute fatal intoxication caused by the pesticide.•A high relative concentration of brain methamidophos may contribute to the intoxication of acephate in humans. We herein report an intoxication case caused by the ingestion of the pesticide Ortoran®, which consists of 50% acephate aqueous solution. A man in his 60 s was found dead in his car with a 100-mL bottle containing approximately 50 mL of Ortoran®. In a gas chromatography – mass spectrometry (GC–MS) screening test, acephate and its metabolite methamidophos were qualitatively detected in his stomach contents. The amounts of acephate and methamidophos (μg/g) in the extract of each body fluid or organ tissue were measured using GC–MS and were as follows: 35.8, 2.84 (heart blood); 44.0, 2.26 (peripheral blood); 2,240, 2.79 (urine); 53.1, 8.91 (brain occipital lobe); 43.7, 2.95 (liver); 102.3, 8.02 (right kidney); and 5450, 22.9 (stomach contents). Based on these results and autopsy findings, the cause of death was concluded to be acute fatal intoxication caused by the pesticide containing acephate and its active metabolite, methamidophos. Concentration ratios between acephate and methamidophos in each body fluid and organ tissue showed higher relative concentrations of brain methamidophos to acephate than those of other organ tissues. A high relative concentration of brain methamidophos may contribute to the intoxication of acephate in humans.

Immunohistochemical investigation of aquaporin (AQP)1 and AQP3 was performed in human skin wounds obtained from forensic autopsy cases. A total of 55 human skin wounds of different postinfliction intervals were collected as follows: group I, 0–3 days (n = 16); II, 4–7 days (n = 11); III, 9–14 days (n = 16); and IV, 17–21 days (n = 12). In uninjured skin samples, AQP1 and AQP3 could be slightly detected in dermal vessels and keratinocytes, respectively. The percentage of AQP1+ vessels and the number of AQP3+ keratinocytes were apparently elevated in accordance with wound ages. The number of AQP3+ keratinocytes was distinctly evident in groups II and III. Morphometrically, both AQP1+ vessel area and AQP3+ cell number were markedly increased in group II, compared with other three groups. With regard to forensic safety, AQP1+ vessel area of over 5% would imply wound ages of 4–12 days. Moreover, the positive area of > 15% would suggest wound age of 7–10 days. Especially, most samples of skin wounds aged 5–10 days except for only one sample (a 10-day-old wound) showed AQP3+ cell number of > 300, and the remaining other samples had that of < 300. Thus, the AQP3+ cell number of > 300 would indicate wound ages of 5–10 days. Collectively, immunohistochemical analyses of AQP1 and AQP3 in human skin wounds would support the objective accuracy of wound age determination.

Endothelial progenitor cells (EPCs), a newly identified cell type, are bone marrow-derived progenitor cells that co-express stem cell markers and vascular endothelial growth factor (VEGF) receptor (Flk-1). In this study, a double-color immunofluorescence analysis was carried out using anti-CD34 and anti-Flk-1 antibodies to examine the time-dependent appearance of EPCs, using 52 human skin wounds with different wound ages (Group I, 0–1 days; Group II, 2–6 days; Group III, 7–14 days; and Group IV, 17–21 days). In wound specimens with an age of less than one day, CD34 + /Flk-1 + EPCs were not detected. EPCs were initially observed in wounds aged two days, and their number was increased in lesions with advances in wound age. In morphometrical analysis, the average number of EPCs was the highest in the wounds of Group III. Especially, 20 out of 21 wounds aged 7–12 days had >20 EPCs, and all wound samples with postinfliction intervals of 14–21 days had <15 EPCs. These observations at least showed that >20 EPCs would indicate a wound age of 7–12 days. Taken together, our observations indicate the detection of EPCs would be useful for wound age determination.

Detection of vitality of mechanical wounds in human cadavers is one of the important issues in forensic medicine. In order to explore novel markers for vitality of acute mechanical wounds, we investigated autophagy in mouse and human skin wounds. Western blotting analysis of mouse skin wounds showed marked reduction of LC3-II and reciprocal increase of p62 in wound samples with the postinfliction intervals of ≥0.5 h, compared with the uninjured skin tissues. These observations indicated that autophagy level was reduced in the wound sites. In postmortem wound samples, there were no remarkable changes in LC3-II and p62 levels. Furthermore, the postmortem intervals of 1–4 days have no significant effects on the changes of LC3-II and p62 in the antemortem skin wounds. Like murine wound samples, these alterations of LC3-II and p62 could be detected in human skin wound samples. Collectively, our study using animal and human samples implied that the detection of autophagy-related molecules such as LC3-II and p62 might be useful for forensic practice as markers of wound vitality.

We investigated the distribution of zolpidem throughout femoral vein blood, urine, stomach contents, the brain, lung, liver, and kidney in five autopsy cases using gas chromatography-mass spectrometry. Four out of the five cases showed toxic levels of zolpidem. Only one case showed a lethal blood level. The zolpidem concentrations in solid organ tissues were generally several times higher than that in blood. Among four solid organ tissues, the differences in concentration were not drastic; they were similar or within the same order of magnitude except for one case. These results suggested that measurements of zolpidem concentrations in multiple solid organ tissues are useful for estimating whether zolpidem poisoning is related to the cause of death, especially when blood cannot be collected.

Fibrocytes, a newly identified cell type, are bone marrow-derived mesenchymal progenitors that coexpress hematopoietic cell antigens and fibroblast products. In this study, a double-color immunofluorescence analysis was carried out using anti-CD45 and anti-collagen type I antibodies to examine the time-dependent appearance of fibrocytes, using 53 human skin wounds with different wound ages (group I, 0–3 days; group II, 4–7 days; group III, 9–14 days; and group IV, 17–21 days). In wound specimens with an age of less than 3 days, CD45 + /collagen type I + fibrocytes were not detected. The fibrocytes were initially observed in wounds aged 4 days, and their number increased in lesions with advances in wound age. In a semiquantitative morphometrical analysis, the average number of fibrocytes was highest in the wounds of group III. These findings imply that human skin wounds containing fibrocytes are at least 4 days old. Moreover, a fibrocyte number of over 10 indicates a wound age between 9 and 14 days (i.e., group III). Based on the average number of fibrocytes in each group, a fibrocyte number of over 15 more strongly suggests a wound age of 9–14 days. Together, our observations indicate the participation of fibrocytes in wound healing of human skin inducing the accumulation of extracellular matrix components, and therefore, detection of fibrocytes could be a useful marker for wound age determination.

Immunohistochemical study combined with morphometry was carried out to examine the expression of cyclooxygenase-2 (COX-2) using 60 human skin wounds of different ages: group I, 0–4 h ( n =  11); II, 8 h–2 days ( n =  21); III, 3–9 days ( n =  14); and IV, 12–21 days ( n =  14). In wound specimens aged 2 h to 2 days, anti-myeloperoxidase-positive neutrophils observed at the wound site expressed immunopositive reaction to COX-2. In wound specimens of more than 3 days, CD68-positive macrophages as well as neutrophils were positively immunostained with anti-COX-2. In group II, all 21 wound samples had COX-2-positive ratios of >40 %, and 15 out of them showed >50 %. In group III, only three wound samples with the postinfliction intervals of 3 days showed positive ratios of 40–50 % and the remaining 11 cases less than 40 %. In groups I and IV, all 25 wound specimens had COX-2-positive ratio of <40 %. With regard to the practical applicability with forensic safety, these observations suggested that a COX-2-positive ratio of >40 % indicated a wound age of 8 h to 3 days. Moreover, COX-2-positive ratios, considerably exceeding a ratio of 50 %, indicate a wound age of 8 h to 2 days. Collectively, COX-2 would be a useful marker for the determination of early wound age.

Triazolam (Trz) concentrations in body fluids and organ tissues of eight autopsy cases were determined by gas chromatography-mass spectrometry (GC-MS). Blood Trz concentrations in seven out of eight cases ranged from less than 2 to 274 ng/g. The distribution of Trz in various organ tissues could be demonstrated in four cases. These distribution data are useful for cadavers that lack blood in forensic autopsy cases. In addition, urinary levels of 1-hydroxymethyltriazolam and 4-hydroxytriazolam, the main metabolites of Trz, were also determined with and without β-glucuronidase treatment by GC-MS for the eight cases. Using the ratios of urine Trz concentration to blood Trz concentration and the ratios of urine 1-hydroxymethyltriazolam concentration to urine 4-hydrozytriazolam concentration, we proposed a method for estimation of intervals between Trz intake and death, although further studies are needed to corroborate it.