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Hepatic macrophages maintain liver homeostasis, but little is known about the signals that activate the hepatoprotective programs within macrophages. Here, we show that toll-like receptor 13 (TLR13), a sensor of bacterial 23S ribosomal RNA (rRNA), senses microbiome RNAs to drive tissue-protective responses in CD5Lhihepatic macrophages. Splenomegaly and hepatomegaly developed in the absence of the endosomal RNase, RNaseT2, via TLR13-dependent macrophage proliferation. Furthermore, TLR13 in hepatic Ly6Clomacrophages activated the transcription factors LXRα and MafB, leading to expression of tissue-clearance molecules, such as CD5L, C1qb, and Axl. Consequently,Rnaset2−/−mice developed resistance to acute liver injury caused by challenges with acetaminophen and lipopolysaccharide + D-galactosamine. TLR13 responses inRnaset2−/−mice were impaired by antibiotics, suggesting that TLR13 were activated by microbiome rRNAs, which was detected in the sera and hepatic macrophages. Repeated administration of wild-type mice with the TLR13 ligand, rather than other TLR ligands, selectively increased the number of Kupffer cells, which expressed immunoregulatory and tissue-clearance genes as hepatic macrophages inRnaset2−/−mice did. Our results suggest that microbiome ssRNA serves as an environmental cue for initiating tissue-protective TLR13 responses in hepatic macrophages.Graphical AbstractIn the absence of an endosomal RNase, RNase T2, microbiome RNAs circulating in the vasculature activate TLR13 in hepatic macrophages to drive hepatoprotective responses through expression of immunoregulatory and tissue-clearance molecules. Consequently, mice lacking RNase T2 are resistant against acute liver injuries caused by acetaminophen and LPS + D-galactosamine.

Supplementary Figure 3 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Supplementary Figure 1 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Accumulating evidence indicates the crucial contribution of chronic inflammation to various types of carcinogenesis, including colon carcinoma associated with ulcerative colitis and asbestosis-induced malignant mesothelioma. Ulcerative colitis–associated colon carcinogenesis can be recapitulated in mice by azoxymethane administration followed by repetitive dextran sulfate sodium ingestion. In the course of this carcinogenesis process, the expression of a macrophage-tropic chemokine, CCL2, was enhanced together with intracolonic massive infiltration of macrophages, which were a major source of cyclooxygenase (COX)-2, a crucial mediator of colon carcinogenesis. Mice deficient in CCL2-specific receptor, CCR2, exhibited less macrophage infiltration and lower tumor numbers with attenuated COX-2 expression. Moreover, CCL2 antagonists decreased intracolonic macrophage infiltration and COX-2 expression, attenuated neovascularization, and eventually reduced the numbers and size of colon tumors, even when given after multiple colon tumors have developed. These observations identify CCL2 as a crucial mediator of the initiation and progression of chronic colitis–associated colon carcinogenesis and suggest that targeting CCL2 may be useful in treating colon cancers, particularly those associated with chronic inflammation. [Cancer Res 2009;69(19):7884–92]

Supplementary Figure 5 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Supplementary Figure 2 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Supplementary Table 2 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Supplementary Figure 4 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Supplementary Figure Legends 1-7 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Supplementary Figure 6 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Supplementary Table 1 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

Supplementary Figure 7 from Blockade of a Chemokine, CCL2, Reduces Chronic Colitis-Associated Carcinogenesis in Mice

In a previous work, authors have proposed a medico-legal definition of femicide as the murder due to the failure to recognize the right of self-determination of women. The aim of this paper was to apply the proposed definition to a cohort of cases to characterise femicides and female homicides and assess whether femicides can be considered a distinct entity or not. A comparison between female and male homicides was performed to assess common and distinctive features. Femicides were identified and compared to the cohort of non-femicide female murder. Results were compared to those reported in published forensic studies. Significant associations between female and male homicides were found for sex and partner/ex-partner offender, sex and indoor homicide and sex and asphyxia as dynamic of death emerged. A higher prevalence of indoor homicides and asphyxiation and of partner relationships were documented in female homicides. Gunshot, blunt injuries and cut wounds are well represented in both types of homicides. Most affected sites are back and chest in male homicides, and head, breasts, pubis, and limbs in female homicides. When comparing femicides and female homicides, a positive association between strangulation as harmful mean and a negative one between femicides and indoor homicides were found. Male and female homicides can be considered as two distinct victimological phenomena. Focusing on femicide allows to establish injuries and circumstantial patterns, that could represent evidence of a specific murder. More studies with a standardized data collection are needed to corroborate the theory of this paper.

Estimating time of death is one of the most important issues in forensic practice. Here, we assessed the applicability, limitation and reliability of a biological clock-based method developed by us. We analyzed the expression of clock genes, BMAL1 and NR1D1, in the heart of the deceased in 318 cases with a defined time of death by real-time RT-PCR. For estimating time of death, we selected two parameters, the NR1D1/ BMAL1 ratio and BMAL1/NR1D1 ratio for death in the morning and evening, respectively. The NR1D1/ BMAL1 ratio was significantly higher in the cases of morning death, while the BMAL1/NR1D1 ratio was significantly higher in the cases of evening death. Gender, age, postmortem interval and most causes of death did not significantly affect the two parameters, except in infant and elderly cases as well as those with severe brain injury. We examined the effects of water intoxication-induced cerebral edema on clock gene expression in mouse hearts and found that cerebral edema alters clock gene expression in this tissue. Based on our findings, we concluded that our method can be applied to forensic practice. However, the method should be applied with caution in infant and elderly cases as well as those with severe brain injury.

Background It has been widely documented that circannual variation has an impact on the incidence and prognosis of cardiovascular diseases. It is unclear why cold ambient temperature increase the incidence of acute myocardial infarction (AMI). We investigated the relationship between the ambient temperature at the onset of AMI, the morphology of the culprit lesion in patients with AMI. Methods We investigated 202 consecutive patients with AMI who underwent optical coherence tomography (OCT). The participants were divided into lower (n = 100) and higher (n = 102) temperature groups based on the ambient temperature. The culprit lesion morphology was compared between the two groups. Results The median temperature at the onset of AMI was 16.6 °C. The prevalence of plaque ruptures was higher at lower temperatures (lower 66% vs. higher 45%, p = .003), whereas OCT-erosion was more frequent in the higher temperature group (lower 13% vs. higher 26%, p = .021). The lower temperature group showed more cholesterol crystals (lower 71% vs. higher 54%, p = .014). Conclusion The peak incidence of AMI in the winter is formed by increased plaque rupture, suggesting environmental temperature has an influence on the pathogenesis of AMI.

This retrospective study evaluated 637 forensic autopsy cases of solitary death (kodoku-shi in Japanese) and clarified the present situation of solitary death from the viewpoints of forensic medicine. Of 637 cases who had solitary death, 471 were male and 166 were female, indicating that solitary death predominantly occurred in men. This tendency increased from the age of 40 years in men and 60 years in women. Among the cases, 349 and 199 were natural death and external death, respectively. Ischemic heart disease- and fire-related fatalities were most common in natural death and external death cases, respectively. Suicidal death was more frequent in women than in men. We found two peaks of postmortem interval until finding (PMI-f): < 1 d (182 cases, 28.6%) and 7 days to < 1 month (164 cases, 25.7%). The PMI-f in female cases was significantly shorter than that in male cases. The PMI-f of external death cases was significantly shorter than that of natural death cases. Kushimoto and Shingu, located in the southern part (Kinan region), had a high solitary death rate in forensic autopsy cases because of their small population and high aging rate. Few cases were welfare recipients in solitary death cases in Kinan region (non-urban area), and 23 of 36 welfare recipients lived in Wakayama City, indicating that the economic state would contribute more to the occurrence of solitary death in Wakayama City (urban area). We identified several risk factors and regional characteristics for solitary death cases, and our observations could contribute to administrative welfare measures for the prevention of solitary deaths.

Inflammatory mediators such as cytokines and chemokines are crucially involved in the development of abdominal aortic aneurysm (AAA). Here we report that CaCl2 application into abdominal aorta induces AAA with intra-aortic infiltration of macrophages as well as enhanced expression of chemokine (C-C motif) ligand 3 (CCL3) and MMP-9. Moreover, infiltrating macrophages express C-C chemokine receptor 5 (CCR5, a specific receptor for CCL3) and MMP-9. Both Ccl3−/− mice and Ccr5−/− but not Ccr1−/− mice exhibit exaggerated CaCl2-inducced AAA with augmented macrophage infiltration and MMP-9 expression. Similar observations are also obtained on an angiotensin II-induced AAA model. Immunoneutralization of CCL3 mimics the phenotypes observed in CaCl2-treated Ccl3−/− mice. On the contrary, CCL3 treatment attenuates CaCl2-induced AAA in both wild-type and Ccl3−/− mice. Consistently, we find that the CCL3–CCR5 axis suppresses PMA-induced enhancement of MMP-9 expression in macrophages. Thus, CCL3 can be effective to prevent the development of CaCl2-induced AAA by suppressing MMP-9 expression., Inflammatory cytokines and chemokines are involved in the development of abdominal aortic aneurysm (AAA). Here the authors show that CCL3 prevents the development of CaCl2-induced AAA by suppressing MMP-9 expression.

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 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.

Dendritic cells (DCs) can essentially contribute to innate and adaptive immune system in various organs. A double-color immunofluorescence analysis was carried out with anti-CD11c and -HLA-DRα antibodies to detect DCs in 53 skin wounds (their postinfliction intervals: group I, 0-3 days; group II, 4-7 days; group III, 9-14 days; and group IV, 17-21 days). CD11c+HLA-DRα+ DCs were first observed in skin wounds with postinfliction intervals of 3 days, and the DC numbers were found to be elevated in skin wounds with the subsequent increase in postinfliction intervals. Semi-quantitative morphometric analyses showed that the DC number was the highest in the 12-day-old wound. More than 50 DCs were present in 8 of 10 samples (80%) in group II and 14 of 16 samples (87.5%) in group III, and there was no difference between the two groups. Thus, the presence of DCs in a skin wound was possibly estimated as postinfliction intervals of at least 3 days. Furthermore, when a skin wound contained > 50 DCs, its age would be judged as 4-14 days. Collectively, the appearance of DCs in human skin wounds may provide useful information in determining the age of a wound.

Graphical abstract, Highlight • The number of in vivo cholesterol crystals at the culprit plaque is increased in patients with the no-reflow phenomenon. • The number of cholesterol crystals, lipid arch, and ostium lesion are independent predictors for the no-reflow phenomenon after PCI. • The combination of the number of cholesterol crystals and lipid arc can improve the prediction ability for the no-reflow phenomenon., Background The release of lipid-laden plaque material subsequent to ST-segment elevation myocardial infarction (STEMI) may contribute to the no-reflow phenomenon. The aim of this study was to investigate the association between in vivo cholesterol crystals (CCs) detected by optical coherence tomography (OCT) and the no-reflow phenomenon after successful percutaneous coronary intervention (PCI) in patients with acute STEMI. Methods We investigated 182 patients with STEMI. Based on the thrombolysis in myocardial infarction (TIMI) flow grade after PCI, patients were divided into a no-reflow group (n = 31) and a reflow group (n = 151). On OCT, CCs were defined as thin, high-signal intensity regions within a plaque. A multivariable logistic regression analysis was performed to determine predictors for the no-reflow phenomenon. Results The prevalence of CCs was higher in the no-reflow group than the reflow group (no-reflow group, 77% vs. reflow group, 53%; p = 0.012). The multivariable logistic model showed that the CC number, lipid arc and ostial lesions were positive independent predictors of no-reflow. The combination of a lipid arc ≥ 139°and CC number ≥ 12 showed good predictive performance for the no-reflow phenomenon (sensitivity, 48%; specificity, 93%; and accuracy, 86%). Conclusion In vivo CCs at the culprit plaque are associated with the no-reflow phenomenon after PCI in patients with STEMI. The combination of the number of CCs and lipid arc can predict the no-reflow phenomenon after PCI with a high accuracy of 86%.

Herein, we report a fatal case of high-pressure water-inflicted femoral injury. A male worker in his twenties was found at the construction site with significant bleeding from the left femoral region. At three hours after discovery, his death was confirmed in an emergency hospital. The deceased was noted to be 182 cm in height, 62.5 kg in weight, and postmortem rigidity strongly appeared in his whole joints. Externally, there was a large, 28-cm laceration in the left medial femoral region, wherein the subcutaneous muscle layer was drastically contused and transected, and both the femoral artery and vein were completely disrupted. This injury also penetrated into the left popliteal region similar to an impalement injury, producing a small, circular, 1.5-cm wound exit site. Moreover, two lacerations in the right anterior femoral region were presented in an inverse “U”-like shape, with injury lengths of 14 cm and 26 cm. Internally, every organ was apparently anemic, given that the water jet lance produced pressures ranging from 10,000–40,000 psi. Thus, the cause of his death was diagnosed as hemorrhagic shock secondary to femoral artery and vein disruption caused by a high-pressure water jet unit.

We immunohistochemically examined the intrathrombotic dynamics of autophagy during thrombogenesis using murine deep vein thrombosis (DVT) models. To perform the immunohistochemical analyses, we used anti-LC3 antibody and anti-p62 antibody for detecting the intrathrombotic autophagic functions. We estimated dynamics of the intrathrombotic autophagy as LC3+ cell number (×1000, five fields) with the thrombus ages (each group n = 5). The number of LC3+ cells was once decreased at 3 days, and then increased until 10 days. On the contrary, the number of p62+ cells progressively increased until 10 days after the inferior vena cava (IVC) ligation, and then gradually decreased. Especially, in all of thrombus samples with the postligation intervals of 5-10 days, both numbers were larger than 10. Subsequently, we compared the number of LC3+ cells to that of p62+ cells. Although, at 1 day after the IVC ligation, LC3+ cell number significantly exceeded p62+ cell number, the former was significantly or relatively less than the latter at 3 days or more after the IVC ligation. Thus, positive cells of > 10 in both LC3 and p62 indicated the thrombus age of 5-10 days. Upon comparison of immunopositive cells in LC3 and p62, the p62/LC3 ratio was > 1.0 in 29 out of 30 thrombus samples aged 3-21 days, and all of 1-day-old thrombus had the p62/LC3 ratio of 1.0 and that of < 0.5 could indicate thrombus age of 3 days or more and that of 1 day, respectively. Collectively, our study implied that the detection of autophagy-related molecules such as LC3 and p62 would be useful for the determination of thrombus age.

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.

Programmed cell death ligand 1 (PD‐L1) on tumor cells suppresses anti‐tumor immunity and has an unfavorable prognostic impact in ovarian cancer patients. We herein report the pathophysiological and therapeutic impacts of PD‐L1 disruption in ovarian cancer. PD‐L1 was genetically disrupted in the murine ovarian cancer cell line ID8 using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9‐mediated genome editing. PD‐L1 knockout (KO) and control ovarian cancer cells were intraperitoneally inoculated into syngeneic mice, and survival and tumor dissemination were evaluated. Survival times were significantly longer in the PD‐L1‐KO ID8‐inoculated groups than in their control groups, and its therapeutic benefit was enhanced in combination with the cisplatin treatment. Tumor weights and ascites volumes were significantly lower in the PD‐L1‐KO ID8 groups than in their control groups. Immunohistochemical and immunofluorescence analyses showed that intratumoral CD4+ T cells, CD8+ T cells, NK cells and CD11c+ M1 macrophages were significantly increased, whereas regulatory T cells were significantly decreased in the PD‐L1‐KO ID8 groups compared with those in their control groups. The intratumoral mRNA expression of interferon‐γ, tumor‐necrosis factor‐α, interleukin (IL)‐2, IL‐12a, CXCL9 and CXCL10 was significantly stronger, while that of IL‐10, vascular endothelial growth factor, CXCL1 and CXCL2 was significantly weaker in the PD‐L1‐KO ID8 groups. These results indicate that CRISPR/Cas9‐mediated PD‐L1 disruption on tumor cells promotes anti‐tumor immunity by increasing tumor‐infiltrating lymphocytes and modulating cytokine/chemokine profiles within the tumor microenvironment, thereby suppressing ovarian cancer progression. These results suggest that PD‐L1‐targeted therapy by genome editing may be a novel therapeutic strategy for ovarian cancer.

Japan is surrounded by the sea and is also a mountainous country with many rivers. Japan has the second- highest rate of deaths caused by drowning in the world. Pleural effusion (PE) is one of the major findings at autopsy. It is found in approximately 80% of drowning mortalities and is observable for a relatively long postmortem interval (PMI). We focused on the amount of pleural fluid in drowning cases, discussed the relationship of PE with the drowning environment, water temperature, and postmortem interval, and established more simple and practical criteria for the diagnosis of drowning. We measured the weight of the lungs, PE, and their sum as the intrathoracic (IT) weight (total weight of lungs + pleural effusion), and calculated the PE ratio [(PE weight/IT weight) × 100]. A total of 130 drowning deaths diagnosed through forensic autopsies were investigated in this study. The cases were classified by drowning environment (freshwater, brackish water, and seawater), water temperature (under 20 °C, more than 20 °C), and postmortem interval (less than 1 day, 1–3 days, more than 3 days). The present study demonstrated that the PE ratio may be more effective for the diagnosis of drowning. Moreover, the accumulation of PE is affected by drowning environment, water temperature, and PMI. Collectively, it is important to assess the PE ratio and consider these factors in autopsy cases of victims found in water.

Asymptomatic patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection must be quickly identified and isolated to prevent the spread of the virus. The number of asymptomatic healthy people is completely unknown because they remain untested. Detection of specific SARS-CoV-2 antibodies has been widely accepted as a diagnostic test, and an immunochromatographic test, which is simpler and relatively cheaper than other methods, is becoming the gold standard for identifying healthy people who had been infected with SARS-CoV-2 in the past. In this study, 1,528 volunteers who worked at a particular hospital were subjected to an immunochromatographic IgG test for SARS-CoV-2 to determine the ratio of asymptomatic people. Only 12 volunteers (0.79%) were IgG+, with no significant background differences in the sex, age, profession, experiences of working at the emergency department or caring for coronavirus disease 2019 patients. If this IgG+ ratio was to be extrapolated to Wakayama city’s population, 2,780 out of 3,54,063 people may be asymptomatic for SARS-CoV-2. The results imply that anyone may get infected with SARS-CoV-2 but remain asymptomatic.