Determining whether ligature marks represent antemortem or postmortem injuries is a critical issue in forensic practice because this distinction directly influences the reconstruction of perimortem events. Consequently, assessing the vital reaction of neck skin plays an essential role in evaluating the viability of compression-related injuries. Cathelicidin antimicrobial peptide (CAMP), a multifunctional host-defense peptide involved in inflammation, immune cell recruitment, and wound repair, may serve as a reliable molecular marker for wound vitality. In this study, we examined CAMP expression in 40 ligature mark samples (32 samples of hanging and 8 samples of ligature/manual strangulation) obtained during forensic autopsies, along with matched intact skin controls, all with postmortem intervals of ≤ 3 days. Immunohistochemistry revealed strong CAMP expression in epidermal keratinocytes and moderate expression in dermal cells at compression sites, whereas control skin showed only faint or absent staining. Morphometric analysis demonstrated that the ratio of CAMP‑positive keratinocytes and the number of CAMP-positive dermal cells were significantly higher in compressed skin than in controls, independent of age, sex or postmortem interval. Moreover, neither ligature type nor mode of neck compression also had no influence on CAMP expression in the keratinocytes or dermal cells. These findings indicate that CAMP expression reflects a biological response to antemortem neck compression and may provide an objective molecular marker for assessing wound vitality in forensic pathology. Furthermore, combining CAMP with other established biomarkers may enhance diagnostic reliability when evaluating antemortem neck compression injuries.

We report a case of fatal acute butane poisoning in a woman in her 80 s, who was found in a bathtub filled with butane gas. Five artificially punctured liquefied petroleum gas (LPG) cartridges were discovered within the bathtub. Forensic autopsy revealed no significant traumatic injuries or pathological findings sufficient to explain the cause of death. Toxicological analysis identified butane and diphenhydramine (DPH) in the blood, urine, and various organs. The peripheral blood concentrations of butane and DPH were 2.95 μ g/ml and 3.56 μ g/ml, respectively. While the DPH concentration was within the toxic range, it did not reach fatal levels independently. Given that the urinary DPH concentration (0.25 μ g/ml) was approximately 1/14th of the blood concentration, it was concluded that death occurred due to butane toxicity during the absorptive phase of DPH. Based on the toxicological findings and the scene circumstances, the cause of death was determined to be acute butane poisoning.

We report a case of sudden and unexpected death due to diabetic ketoacidosis in a severely obese young woman (height, 146 cm; weight, 98 kg) with no prior diagnosis of diabetes mellitus but a history of prolonged excessive consumption of sugary beverages. Autopsy and histopathological findings revealed no fatal organic pathology except for severe fatty liver, and the toxicological examination was negative. Postmortem biochemical analysis showed markedly elevated serum glucose (1890 mg/dL), vitreous glucose (1900 mg/dL), glycated hemoglobin (13.7

We herein report a case of sudden death from massive mucinous cystadenoma of the ovary. A female in her 70s was found dead with an enlarged abdomen. Forensic autopsy was performed to identify the cause of death. Postmortem computed tomography showed a large amount of intra-abdominal fluid pushing against intraperitoneal organs. The deceased was 152 cm in height and weighed 85 kg. The abdomen was enlarged with a girth of 133 cm. A left ovarian cyst was detected and contained 32 L of a dark brown fluid. The uterus and ovaries weighed 1200 g. The heart weighed 400 g, and atherosclerosis was noted in the left coronary artery and aorta. The pathophysiological mechanisms leading to death were considered to be multifactorial. The massive ovarian cyst increased intra-abdominal pressure and produced a cephalic shift in the diaphragm. This may have had a negative impact on the cardiovascular and respiratory systems by (a) increasing the heart rate, (b) decreasing lung compliance, (c) increasing the respiratory rate, (d) increasing maximum inspiratory pressure, (e) increasing mean arterial pressure and systemic vascular resistance, (f) reducing femoral venous return, (g) compressing the inferior vena cava, and (h) decreasing venous return. This, in turn, increased susceptibility to hypoxia in addition to the underlying ischemic heart disease. We diagnosed the cause of death as acute circulation failure due to myocardial ischemia based on intra-abdominal hypertension as a result of the massive ovarian cyst.

This retrospective study evaluated 637 forensic autopsy cases of solitary deaths (kodoku-shi in Japanese) and clarified the current status of solitary deaths from the viewpoint of forensic medicine. Of the 637 cases of solitary death, 471 were men and 166 were women, indicating that solitary death predominantly occurred in men. This tendency increased from the age of 40 years in men to 60 years in women. Among these, 349 and 199 were natural and external deaths, respectively. Ischemic heart disease and fire-related fatalities were the most common causes of natural and external death, respectively. Suicidal deaths were more frequent among women than men. We found two peaks of postmortem interval until finding (PMI-f): <1 day (182 cases, 28.6 %) and 7 days to < 1 month (164 cases, 25.7 %). The PMI-f was significantly shorter in women than in men. The PMI-f of external death cases was significantly shorter than that of natural death cases. Kushimoto and Shingu, located in Southern Japan (Kinan region), had a high solitary death rate in forensic autopsy cases because of the high aging rate and high rate of individuals in single-person households aged ≥ 65 years. Few cases of solitary deaths in the Kinan region (non-urban area) were welfare recipients, and 23 of the 36 welfare recipients lived in Wakayama City, indicating that the economic state contributed more to the occurrence of solitary deaths in Wakayama City (urban area). We identified several risk factors and regional characteristics of solitary deaths. Our observations could contribute to administrative welfare measures for the prevention of solitary deaths.

We herein describe a suicidal death due to the excessive ingestion of over-the-counter caffeine pills. A female in her 20 s with psychiatric disorders was found dead at her home. A crime scene investigation by police revealed 18 empty sheets, corresponding to ca. 20 g of caffeine, in her room. Forensic autopsy showed that there were neither lethal injuries nor pathological lesions in any organ. In the stomach, there was dark brown-colored food debris weighing 180 g without any tablets. Toxicological analyses revealed that caffeine concentrations in intracardiac and peripheral blood samples reached fatal levels. Therefore, the cause of death was diagnosed as acute caffeine intoxication. To the best of our knowledge, autopsy case reports from caffeine intoxication are rare; however, the risk of caffeine intoxication has increased due to the easy availability of caffeine products. This case report compels governments to restrict the purchase of caffeine at drugstores or through e-markets.

Japanese spotted fever (JSF) is potentially fatal infection transmitted by tick bites which vectors Rickettsia (R.) japonica. Since JSF was first described in 1984, the incidence has gradually been increased. We experienced a case of JSF of fatal outcome. A female in 70 ' s was found dead on her bed, whose house was so called 'hoarding house' filled with many waists and unused items. The following day, the autopsy was performed. As representative symptom of external findings, skin rashes were seen on the trunk and extremities, and there were tick-bite eschars on the left upper arm. Internal findings showed no specific findings in each organ. Histopathological examination demonstrated massive inflammatory cell infiltrates mainly consisted of neutrophils in the dermis beneath the external eschar. Furthermore, destruction of glomeruli in kidney with microhemorrhage from mesangial regions was observed. The numerous inflammatory infiltrates were also observed in pulmonary interstitium, which were accompanied with histopathologic features of vasculitis. Biochemical examination showed severe systemic inflammation as monitored by elevated CRP of 16 mg/dL and renal dysfunction by BUN of 171.2 mg/dL and creatinine of 6.07 mg/dL. Subsequently polymerase chain reaction revealed specifically amplified signals for R. japonica from the samples of tick-bites eschar and blood. Thus, we diagnosed her cause of death as JSF which had been occurred multiorgan failure such as acute renal failure and possibly acute respiratory failure. (224 terms)

When exposed to oxidative and electrophilic stress, a protective antioxidant response is initiated by nuclear factor erythroid 2-related factor 2 (Nrf2). However, the extent of its importance in the forensic diagnosis of acute ischemic heart diseases (AIHD), such as myocardial infarction (MI), remains uncertain. On the other hand, immunohistochemical analyses of fibronectin (FN) and the terminal complement complex (C5b-9) prove valuable in identifying myocardial ischemia that precedes necrosis during the postmortem diagnosis of sudden cardiac death (SCD). In this study, we investigated the immunohistochemical levels of Nrf2, FN, and C5b-9 in human cardiac samples to explore their forensic relevance for the identification of acute cardiac ischemia. Heart samples were obtained from 25 AIHD cases and 39 non-AIHD cases as controls. Nrf2 was localized in the nuclei of cardiomyocytes, while FN and C5b-9 were detected in the myocardial cytoplasm. The number of intranuclear Nrf2 positive signals in cardiomyocytes increased in AIHD cases compared to control cases. Additionally, the grading of positive portions of cardiac FN and C5b-9 in the myocardium was also significantly enhanced in AIHD, compared to controls. Collectively, these results indicate that the immunohistochemical investigation of Nrf2 combined with FN, and/or C5b-9 holds the potential for identifying early-stage myocardial ischemic lesions in cases of SCD.

Immunohistochemical analysis of platelet-derived growth factor receptor-α (PDGFR-α) was performed on human skin wounds obtained from forensic autopsy cases. Thirty human skin wounds were collected at different post-infliction intervals as follows: Group I, 4 h to 3 days (n = 16); Group II, 4 to 7 days (n = 7); Group III, 9 to 10 days (n = 3); and Group IV, 14 to 20 days (n = 4). Immunopositive reactions for PDGFR-α were not observed in the uninjured human skin specimens. In a semi-quantitative morphometrical analysis, the number of PDGFR-α-positive cells was observed increased in Group II, with the average number of PDGFR-α-positive cells being the highest in Group II. Additionally, in Group II, all specimens showed PDGFR-α-positive cells, with an average number of > 200 cells in five fields of view, suggesting a wound age of 4 to 7 days. Taken together, the immunohistochemical detection of PDGFR-α in human skin wounds can be a useful tool for wound age determination.

Cisplatin is an effective chemotherapeutic agent widely used for the treatment of various solid tumors. However, cisplatin has an important limitation in its use; currently, there is no method to ameliorate cisplatin-induced acute kidney injury (AKI). Thrombomodulin (TM) is well known not only for its role as a cofactor in the clinically important natural anticoagulation pathway but also for its anti-inflammatory properties. Here, we investigated the effects of TM in cisplatin-induced AKI. In mice intraperitoneally injected with 15 mg/kg cisplatin, TM (10 mg/kg) or PBS was administered intravenously at 24 h after cisplatin injection. TM significantly attenuated cisplatin-induced nephrotoxicity with the suppressed elevation of blood urea nitrogen and serum creatinine, and reduced histological damages. Actually, TM treatment significantly alleviated oxidative stress-induced apoptosis by reducing reactive oxygen species (ROS) levels in cisplatin-treated renal proximal tubular epithelial cells (RPTECs) in vitro. Furthermore, TM clarified cisplatin-induced apoptosis by reducing caspase-3 levels. In addition, TM attenuated the endoplasmic reticulum (ER) stress signaling pathway in both renal tissues and RPTECs to protect the kidneys from cisplatin-induced AKI. These findings suggest that TM is a potential protectant against cisplatin-induced nephrotoxicity through suppressing ROS generation and ER stress in response to cisplatin.

Heat shock proteins (HSPs) are molecular chaperones whose primary function is cytoprotection, supporting cell survival under (sub) lethal conditions. They have been implicated in various diseases such as inflammatory diseases and cancer due to their cytoprotective and immunomodulatory effects, and their biological mechanisms have been studied. Central family members include, HSP27, which is induced by various stimuli such as heat shock, hypoxia, hyperoxia, ultraviolet exposure, and nutritional deficiency, and HSP70, which is homeostatically expressed in many organs such as the gastrointestinal tract and has anti-cell death and anti-inflammatory effects. In this study, HSP27 and HSP70 were investigated during thrombus formation and dissolution in a deep vein thrombosis model by immunohistochemistry to determine their involvement in this process and whether their expression could be used as a forensic marker. In the process of thrombus formation and lysis, HSP27 and HSP70 were found to be expressed by immunohistochemical analysis. The role of inhibitors of HSP27 and HSP70 in the pathogenesis of thrombosis in mice was also investigated. When HSP27 or HSP70 inhibitors were administered, thrombi were significantly smaller than in the control group on day 5 after inferior vena cava ligation, indicating pro-thrombotic effects HSP27 and HSP70. If HSP27- or HSP70-positive cells were clearly visible and easily identifiable in the thrombus sections, the thrombus was presumed to be more than 10 days old. Thus, the detection of intrathrombotic HSP27 and HSP70 could forensically provide useful information for the estimation of thrombus ages. Collectively, our study implied that both HSP27 and HSP70 might be molecular targets for thrombus therapy and that the detection of HSP-related molecules such as HSP27 and HSP70 could be useful for the determination of thrombus ages.

Aquaporins (AQPs) are a family of water channel proteins that primarily elicit the basic functions of water transport and osmotic homeostasis. To date, at least 17 mammalian AQPs have been identified, AQP-0 to -12 have been found in higher orders including human, and AQP-13 to -16 have been described in older lineages. Moreover, these proteins have recently been shown to regulate many biological processes through unique activities, such as cell proliferation, migration, apoptosis, and mitochondrial metabolism. Several studies have focused on the involvement of AQPs in cell biology aspect, showing that they are involved in a variety of physiological processes and pathophysiological conditions. Furthermore, in the field of forensic medicine, studies on whether AQPs can be a useful marker for diagnosing various causes of death have been conducted using autopsy samples and animal experiments, which have produced interesting results. Herein, we review certain observations regarding AQPs and discuss their potential to contribute to the future practice of forensic research.

Intense neutrophil infiltration into the liver is a characteristic of acetaminophen-induced acute liver injury. Neutrophil elastase is released by neutrophils during inflammation. To elucidate the involvement of neutrophil elastase in acetaminophen-induced liver injury, we investigated the efficacy of a potent and specific neutrophil elastase inhibitor, sivelestat, in mice with acetaminophen-induced acute liver injury. Intraperitoneal administration of 750 mg/kg of acetaminophen caused severe liver damage, such as elevated serum transaminase levels, centrilobular hepatic necrosis, and neutrophil infiltration, with approximately 50% mortality in BALB/c mice within 48 h of administration. However, in mice treated with sivelestat 30 min after the acetaminophen challenge, all mice survived, with reduced serum transaminase elevation and diminished hepatic necrosis. In addition, mice treated with sivelestat had reduced NOS-II expression and hepatic neutrophil infiltration after the acetaminophen challenge. Furthermore, treatment with sivelestat at 3 h after the acetaminophen challenge significantly improved survival. These findings indicate a new clinical application for sivelestat in the treatment of acetaminophen-induced liver failure through mechanisms involving the regulation of neutrophil migration and NO production.

Estimating the age and vitality of human skin wounds is essential in forensic practice, and the use of immunohistochemical parameters in this regard remains a challenge. Heat shock proteins (HSPs) are evolutionarily conserved universal proteins that protect biological systems from various types of stress. However, its importance in forensic pathology for determining wound activation in neck compression skin remains unclear. The expression of HSP27 and HSP70 in neck skin samples was immunohistochemically examined to understand its forensic applicability in determining wound vitality. Skin samples were obtained from 45 cases of neck compression (hanging, 32 cases; strangulation, 10 cases; manual strangulation, 2 cases; other, 1 case) during forensic autopsies; intact skin from the same individual was used as a control. HSP27 expression was detected in 17.4% of keratinocytes in the intact skin samples. In the compressed region, the frequency of HSP27 expression in keratinocytes was 75.8%, which was significantly higher than that in intact skin. Similarly, HSP70 expression was 24.8% in intact skin samples and 81.9% in compressed skin samples, significantly higher in compressed skin than in intact skin samples. This increase in case compression cases may be due to the cell defence role of HSPs. From a forensic pathology perspective, the immunohistochemical examination of HSP27 and HSP70 expression in neck skin could be considered a valuable marker for diagnosing traces of antemortem compression.

Estimating time of death is one of the most important problems in forensics. Here, we evaluated the applicability, limitations and reliability of the developed biological clock-based method. We analyzed the expression of the clock genes, BMAL1 and NR1D1, in 318 dead hearts with defined time of death by real-time RT-PCR. For estimating the time of death, we chose two parameters, the NR1D1/BMAL1 ratio and BMAL1/NR1D1 ratio for morning and evening deaths, respectively. The NR1D1/BMAL1 ratio was significantly higher in morning deaths and the BMAL1/NR1D1 ratio was significantly higher in evening deaths. Sex, age, postmortem interval, and most causes of death had no significant effect on the two parameters, except for infants and the elderly, and severe brain injury. Although our method may not work in all cases, our method is useful for forensic practice in that it complements classical methods that are strongly influenced by the environment in which the corpse is placed. However, this method should be applied with caution in infants, the elderly, and patients with severe brain injury.

We report a case of hemoperitoneum after percutaneous radiofrequency ablation in a patient with hepatocellular carcinoma. A 60-year-old female was hospitalized for the treatment of thrombasthenia and cirrhosis caused by chronic Hepatitis C, and computed tomography revealed hepatocellular carcinoma, which was treated by percutaneous radiofrequency ablation. After the ablation, hemoperitoneum was suspected because of the low hemoglobin level with abdominal pain. Approximately 6 h after the ablation treatment, the patient suddenly fell into a shock state and died. In this case, medical treatment-related death including malpractice was suspected, and forensic autopsy was performed. The abdominal cavity contained 910 mL of dark red fluid blood and 210 g of soft hemocoagula. Moreover, several puncture marks were observed on the liver surface and diaphragm, and there was no clear damage to the main arteries and veins. Considering the macroscopic and microscopic findings, the cause of death was assumed as hemorrhagic shock due to the hemoperitoneum caused by the damage to the liver by radiofrequency ablation. It is important to consider all the indications and adverse effects of radiofrequency ablation.

Background Deep vein thrombosis (DVT) is a common peripheral vascular disease arising from endothelial damage and can frequently result in pulmonary embolism. C-C chemokine receptor 5 (CCR5) is essentially involved in skin wound healing and pulmonary fibrosis, but the role of CCR5 still remains elusive. Methods Deep vein thrombus was induced by the ligation of the inferior vena cava (IVC) in wild-type (WT) mice and Ccr5 -/- mice, and thrombi were collected over time and various analyzes were performed. Results After ligation of the IVC in WT mice, a venous thrombus developed progressively until 5 days, and then resolved. Concomitantly, IVC ligation enhanced intrathrombotic gene and protein expression of Ccr5 and its ligand, Ccl5 , and both were expressed mainly by intrathrombotic macrophages. The same treatment of Ccr5 -/- mice resulted in significantly greater thrombus mass than WT mice. Moreover, the administration of a specific CCR5 inhibitor to WT mice recapitulated similar phenotypes as Ccr5 -/- mice while that of CCL5 caused the opposite phenotypes. In Ccr5 -/- mice, the production of intrathrombotic vascular endothelial growth factor (VEGF), tissue plasminogen activator (tPA), and urokinase-type plasminogen activator (uPA) from macrophages was significantly reduced and intrathrombotic recanalization was also suppressed compared to WT mice. Moreover, CCL5 enhanced Vegf, Plat, and Plau gene expression in WT - derived peritoneal macrophages by activating the ERK MAPK signaling pathway in vitro. Conclusions CCR5-deficient mice exhibited reduced expression of Vegf, Plat, and Plau, with concomitant attenuated neovascularization and reduced thrombus resolution. the Thus, the augmentation of the CCL5-CCR5 axis may be effective for DVT treatment by enhancing thrombolysis.

We investigated the dynamics of the gene expression of M1 and M2 macrophage markers during skin wound healing in mice. Expression of M1-macrophage markers, such as Il12a, Tnf, Il6, Il1b, and Nos2 was upregulated after wounding and peaked at 1 or 3 days after injury, and that of M2-macrophage markers such as Mrc1, Cd163, Ccl17, Arg, and Tgfb1, peaked at 6 days after injury. Consistent with these findings, using triple-color immunofluorescence analysis revealed that F4/80 + CD80 + M1 macrophages were more abundant than F4/80 + CD206 + M2 macrophages on day 3 in mouse wound specimens, and that M2 macrophages were prominently detected in day 6 wounds. For application in forensic practice, we examined macrophage polarization using human wound specimens. The average ratios of CD68 + iNOS + M1 macrophages to CD68 + CD163 + M2 macrophages (M1/M2 ratios) were greater than 2.5 for the wounds aged 2–5 days. Out of 11 wounds aged 1–5 days, five samples had the M1/M2 ratios of > 3.0. These observations propose that the M1/M2 ratios of 3.0 would indicate a wound age of 1–5 days as the forensic opinion. This study showed that M1 and M2 macrophages in human skin wound might be a promising marker for wound age determination.

ABSTR A C T Here, we report a case of necrotizing fasciitis following intra-articular injection of hyaluronic acid. A 73-year-old female received intra-articular injections of hyaluronic acid due to arthralgia at the left shoulder and knee, and was found dead in her living room at one day. At the forensic autopsy, injection marks with bullae and erythema were found at the left shoulder and knee and liquefactive necrosis of muscle tissues was observed in the left but not right extremities. Histopathological examinations of the left upper arm and thigh revealed severe rhabdo-myolysis with lots of bacterial clusters. Bacteriological examinations detected group A Streptococcus from intracardiac blood and affected muscle tissues. Postmortem biochemical analysis of blood showed escalated blood urea nitrogen (133.8 mg/dL), creatinine (4.57 mg/dL) and C-reactive protein (45.0 mg/dL). The cause of her death was diagnosed as streptococcal toxic shock syndrome (STSS). Moreover, it was suggested that the injection was inappropriately conducted and served as a portal of bacterial entry.