Endothelial CLEC-1b plays a protective role against cancer hematogenous metastasis.

Metastasis, which is the spread of cancer cells into distant organs, is a critical determinant of prognosis in patients with cancer, and blood vessels are the major route for cancer cells to spread systemically. Extravasation is a critical process for the hematogenous metastasis; however, its underlying molecular mechanisms remain poorly understood. Here, we identified that senescent ECs highly express C-type lectin domain family 1 member B (CLEC-1b), and that endothelial CLEC-1b inhibits the hematogenous metastasis of a certain type of cancer. CLEC-1b expression was enhanced in ECs isolated from aged mice, senescent cultured human ECs, and ECs of aged human. CLEC-1b overexpression in ECs prevented the disruption of endothelial integrity, and inhibited the transendothelial migration of cancer cells expressing podoplanin (PDPN), a ligand for CLEC-1b. Notably, target activation of CLEC-1b in ECs decreased the hematogenous metastasis in the lungs by cancer cells expressing PDPN in mice. Our data reveal the protective role of endothelial CLEC-1b against cancer hematogenous metastasis. Considering the high CLEC-1b expression in senescent ECs, EC senescence may play a beneficial role with respect to the cancer hematogenous metastasis.

Estimation of cadaveric age in crime scenes using Raman spectroscopy.

The number of unidentified corpses has been increasing in recent years. There is a need for an objective and readily applicable method to estimate age, which is important information for identification. In previous reports, we reported that the protein folding ratio (RPF) of skin, as measured by Raman spectroscopy using cross sections of skin samples, is highly correlated with age. In this study, we investigated the possibility of estimating age by measuring Raman spectra from the skin surface of cadavers using a portable device. The resultant intercept, slope, and root mean square error were 97.9, - 63.7 (p < 0.0001), and 11.68, respectively. We evaluated this regression formula by using 10-fold cross-validation, resulting in a coefficient of determination of 0.51. The portable Raman spectrometer may be of assistance in estimating age at death of corpses at the scene of discovery.

Postmortem computed tomography imaging of negative pressure pulmonary edema induced by rice cakes: A case report.

Airway obstruction by foreign bodies or laryngospasms often results in negative pressure pulmonary edema (NPPE). NPPE can develop despite the removal of airway obstruction. This is well-known among clinicians, particularly anesthesiologists, emergency physicians, pediatricians, and radiologists. Computed tomography (CT) features of NPPE include diffuse central ground-glass opacities and thickening of the bronchovascular bundles and bronchial walls. NPPE can also occur after strangulation or smothering with a plastic bag, and is sometimes fatal. Nevertheless, no reports of NPPE are available in forensic journals, and forensic practitioners may not be dedicating the necessary attention to this condition. Recently, the use of postmortem CT as an auxiliary imaging modality for autopsies has increased among forensic institutions in Japan. Understanding the mechanisms and CT findings of NPPE will be useful for forensic practitioners. Herein, we present a case wherein the postmortem CT image revealed NPPE resulting from the obstruction of the airway induced by rice cakes.

Artificial intelligence-based forensic sex determination of East Asian cadavers from skull morphology.

Identification of unknown cadavers is an important task for forensic scientists. Forensic scientists attempt to identify skeletal remains based on factors including age, sex, and dental treatment remains. Forensic scientists commonly consider skull or pelvic shape to evaluate the sex; however, these evaluations require sufficient experience and knowledge and lack objectivity and reproducibility. To ensure objectivity and reproducibility for sex evaluation, we applied a gated attention-based multiple-instance learning model to three-dimensional (3D) skull images reconstructed from postmortem head computed tomography scans. We preprocessed the images, trained with 864 training data, validated the model with 124 validation data, and evaluated the performance of our model in terms of accuracy with 246 test data. Furthermore, three forensic scientists evaluated the 3D skull images, and their performances were compared with those of the model. Our model showed an accuracy of 0.93, which was higher than that of the forensic scientists. Our model primarily focused on the entire skull owing to visualization but focused less on the areas often investigated by forensic scientists. In summary, our model may serve as a supportive tool to identify cadaver sex based on skull shape. Further studies are required to improve the model's performance.

Factors determining the resection ability of snares in cold snare polypectomy: Construction of an ex vivo model for accurately evaluating resection ability.

OBJECTIVES: Cold snare polypectomy (CSP)-dedicated snares (DSs) may have a higher resection ability than conventional snares. However, a model that can accurately and objectively evaluate and compare the resection ability of each snare has yet to be determined, and characteristics of snare parts that increase resection ability remain unknown. Therefore, we elucidated DSs' resection ability and all characteristics of the parts required for acquiring high resection ability. METHODS: An ex vivo model for evaluating resection ability was generated using human colons obtained from forensic autopsy specimens. The force required to resect a 15 mm wide human colonic mucosa (FRR) was measured using this model; if the FRR is small, the resection ability is high. Next, after measuring the stiffness of each snare part, the correlation between the stiffness and resection ability was analyzed. RESULTS: The force required to resect using SnareMaster Plus, Micro-Tech Cold Snare, Captivator Cold, Exacto Cold Snare, or Captivator II was 13.6 ± 1.0, 12.5 ± 1.2, 7.4 ± 1.2, 6.5 ± 1.0, and 28.7 ± 3.7 N, respectively. All DSs had significantly lower FRR than the conventional snare (Captivator II) and had higher resection ability (P < 0.001). A negative correlation was found between FRR and sheath or wire spindle stiffness, with correlation coefficients of 0.72 (P = 0.042) or 0.94 (P < 0.001), respectively. Moreover, 1 × 7 type wire rings had significantly higher friction coefficients than 1 × 3 type wire rings (P < 0.002). CONCLUSION: Sheath and wire spindle stiffness should be increased to increase resection ability; 1 × 7 type wire rings may be suitable for CSP-snare parts.

Detection of human chorionic gonadotropin (HCG) in fabric urine stains older than five years.

Human chorionic gonadotropin (HCG) is used in clinical medicine as a particularly important indicator to determine pregnancy. In this study, it was necessary to determine whether the urine spots on car seat fabric from a murder 5 years previously were from a pregnant woman. The HCG in the dried urine spot on a car seat was detected using an immunochromatography kit. It was found that the HCG in urine can be detected for much longer periods of time than the previously reported period of approximately 6 months.

A suicide case by hanging without fixing the end of the rope.

Hanging is a method of death in which one end of a cord is tied around the neck and the other end is tied to an unmovable object and hung down, compressing the neck with weight. We have identified a rare case in which death was accomplished without tying one end of the rope. The individual was successfully hanged by the frictional force between the upward rope and the downward rope. The police concluded the case as a suicide.

Deep Neural Networks-Based Age Estimation of Cadavers Using CT Imaging of Vertebrae.

Although age estimation upon death is important in the identification of unknown cadavers for forensic scientists, to the best of our knowledge, no study has examined the utility of deep neural network (DNN) models for age estimation among cadavers. We performed a postmortem computed tomography (CT) examination of 1000 and 500 male and female cadavers, respectively. These CT slices were converted into 3-dimensional images, and only the thoracolumbar region was extracted. Eighty percent of them were categorized as training datasets and the others as test datasets for both sexes. We fine-tuned the ResNet152 models using the training datasets. We conducted 4-fold cross-validation, and the mean absolute error (MAE) of the test datasets was calculated using the ensemble learning of four ResNet152 models. Consequently, the MAE of the male and female models was 7.25 and 7.16, respectively. Our study shows that DNN models can be useful tools in the field of forensic medicine.

Evaluation of Environmental Stability and Disinfectant Effectiveness for Human Coronavirus OC43 on Human Skin Surface.

The environmental stability of human coronavirus OC43 (HCoV-OC43) on the surface of human skin and the effectiveness of disinfectant against HCoV-OC43, which are important to prevent contact transmission, have not been clarified in previous studies. Using previously generated models, we evaluated HCoV-OC43 stability and disinfection effectiveness. Then we compared the results with those for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The median survival time of HCoV-OC43 on the surface of human skin was 24.6 h (95% confidence interval, 19.7 to 29.6 h), which was higher than that of SARS-CoV-2 (10.8 h). Although the in vitro disinfectant effectiveness evaluation showed that HCoV-OC43 has a higher ethanol resistance than SARS-CoV-2, HCoV-OC43 on the skin surface was completely inactivated by a minimum of 50% ethanol within 5 s (the log reduction values were >4.0). Moreover, 1.0% chlorhexidine gluconate and 0.2% benzalkonium chloride showed relatively high disinfectant effectiveness, and the log reduction values when these disinfectants were applied for 15 s were >3.0. HCoV-OC43 is highly stable on the skin surface, which may increase the risk of contact transmission. Although HCoV-OC43 has relatively high ethanol resistance, appropriate hand hygiene practices with current alcohol-based disinfectants sufficiently reduce the risk of contact transmission. IMPORTANCE This study revealed the environmental stability of HCoV-OC43 and disinfectant effectiveness against HCoV-OC43, which had not been demonstrated in previous studies. HCoV-OC43 is highly stable on the surface of human skin, with a survival time of approximately 25 h. High stability of HCoV-OC43 may increase the risk of contact transmission. Furthermore, the in vitro disinfectant effectiveness evaluation showed that HCoV-OC43, which is classified as an envelope virus, has a relatively high ethanol resistance. This finding suggests that disinfectant effectiveness may vary greatly depending on the virus and that each virus targeted for infection control should be evaluated individually. HCoV-OC43 on the skin surface was rapidly inactivated by 50% ethanol, which suggests that appropriate hand hygiene practices with current alcohol-based disinfectants can sufficiently reduce the risk of HCoV-OC43 contact transmission.

Decrease in the expression of muscle-specific miRNAs, miR-133a and miR-1, in myoblasts with replicative senescence.

Muscles that are injured or atrophied by aging undergo myogenic regeneration. Although myoblasts play a pivotal role in myogenic regeneration, their function is impaired with aging. MicroRNAs (miRNAs) are also involved in myogenic regeneration. MiRNA (miR)-1 and miR-133a are muscle-specific miRNAs that control the proliferation and differentiation of myoblasts. In this study, we determined whether miR-1 and miR-133a expression in myoblasts is altered with cellular senescence and involved in senescence-impaired myogenic differentiation. C2C12 murine skeletal myoblasts were converted to a replicative senescent state by culturing to a high passage number. Although miR-1 and miR-133a expression was largely induced during myogenic differentiation, expression was suppressed in cells at high passage numbers (passage 10 and/or passage 20). Although the senescent myoblasts exhibited a deterioration of myogenic differentiation, transfection of miR-1 or miR-133a into myoblasts ameliorated cell fusion. Treatment with the glutaminase 1 inhibitor, BPTES, removed senescent cells from C2C12 myoblasts with a high passage number, whereas myotube formation and miR-133a expression was increased. In addition, primary cultured myoblasts prepared from aged C57BL/6J male mice (20 months old) exhibited a decrease in miR-1 and miR-133a levels compared with younger mice (3 months old). The results suggest that replicative senescence suppresses muscle-specific miRNA expression in myoblasts, which contributes to the senescence-related dysfunction of myogenic regeneration.

Application of Teeth in Toxicological Analysis of Decomposed Cadavers Using a Carbamazepine-Administered Rat Model.

In a regular autopsy, blood and organs are used to quantify drug and toxicant concentrations; however, specimens such as blood cannot be collected from highly decomposed corpses, making the quantification of drug and toxicants impossible. This study aimed to estimate the blood carbamazepine (CBZ) concentration from teeth, a part of the human body that is best preserved after death. We sampled teeth and blood of rats administered CBZ. The correlation between the tooth and serum CBZ concentrations was analyzed. Rats were euthanized after CBZ administration and kept at 22 °C for 0 to 15 days before sampling the teeth and measuring the CBZ concentration. Undecalcified, fresh, frozen sections of rat teeth were prepared, and CBZ localization was evaluated. CBZ concentrations in both teeth and cardiac blood peaked at 60 min after administration and increased in a dose-dependent manner. CBZ concentration in teeth did not substantially change after death, with high CBZ distribution being observed in the pulp cavity. The tooth and serum CBZ concentrations were highly correlated, suggesting that the measurement of toxicant concentration in sampled teeth would allow for the estimation of blood toxicant concentration in highly decomposed corpses.

Pentobarbital overdose: A case of contract killing.

Suicides by pentobarbital overdose have increased since about 2012, which appear to be influenced by technical information on active euthanasia that has spread over the Internet. We encountered a pentobarbital poisoning case of a patient with amyotrophic lateral sclerosis. A caregiver found the patient unconscious immediately after two visitors left the room. The patient was immediately transferred to the emergency hospital but eventually declared dead. A fatal concentration of pentobarbital was detected in peripheral blood samples collected in the emergency hospital and during autopsy (53.8 µg/mL and 29.4 µg/mL, respectively). Because the ratios of pentobarbital concentrations between the gastric contents and peripheral blood were 35 and 29 in the hospital and autopsy samples, respectively, it is likely that pentobarbital was administered via the gastrostomy tube. The patient had contacted the visitors through social media. Although the patient had requested the doctor perform active euthanasia and expressed a desire to end their life on social media, nobody had noticed the plan to commit suicide.

Sudden death of a 2-year-old child due to alpha-ketoadipic aciduria.

Alpha-ketoadipic acid is one of the metabolic intermediates of lysine and tryptophan, and it is known as the biochemical hallmark of alpha-ketoadipic aciduria (α-KA). α-KA is a rare autosomal recessive disorder. Its pathophysiology is reduced alpha-ketoadipic acid dehydrogenase activity, and that makes it difficult to metabolize lysine and tryptophan. The symptoms of this disease are multiple, e.g., psychomotor retardation, epilepsy, and ataxia, and it can even be asymptomatic. We present a case of sudden death in a 2-year-old boy with alpha-ketoadipic aciduria. Postmortem computed tomography (CT) and autopsy were performed to elucidate the cause of death. No obvious lesions could be identified except for a marked fatty liver. Urinalysis showed elevated excretion of α-ketoadipic acid.

Estimating the age at death for forensic cases using quantitative computed tomography.

Estimation of the age at death is an important task for forensic scientists. Although the correlation between age and bone mineral density is already known, including for cadavers, to our knowledge, there are no published studies on age estimation with quantitative computed tomography. Quantitative computed tomography can be used to measure bone mineral density based on the mean computed tomography value of the cancellous bone. As this value cannot be calculated in putrefied cases, we modified quantitative computed tomography to calculate the bone mineral density from regions of the bone with mean computed tomography values of 50-350 Hounsfield units. We aimed to examine whether this method could be used for age estimation. We examined 171 male and 106 female cadavers, some of which were putrefied. We performed univariate linear regression analysis for age at death and bone mineral density. The resultant intercept, slope, and root mean square error were 91.3, - 0.20 (p < 0.0001), and 11.4, respectively, for male cadavers, and 96.1, - 0.23 (p < 0.0001), and 11.0, respectively, for female cadavers. We evaluated this regression formula by using 10-fold cross-validation, resulting in a coefficient of determination of 0.33 for male cadavers and 0.42 for female cadavers. The modified quantitative computed tomography method may be of assistance in estimating age at death, even in putrefied cases.

Differences in environmental stability among SARS-CoV-2 variants of concern: both omicron BA.1 and BA.2 have higher stability.

OBJECTIVES: The increased infectivity and transmissibility of SARS-CoV-2 variants of concern (VOCs) could cause significant human and economic damage. Hence, understanding their characteristics is crucial to control infection. We evaluated the environmental stability of the Wuhan strain and all VOCs (Alpha, Beta, Gamma, Delta, Omicron BA.1, and Omicron BA.2 variants) on plastic and human skin surfaces and their disinfection efficacy. METHODS: To evaluate environmental stability, residual virus titres on plastic and human skin surfaces were measured over time. Their survival time and half-life were calculated using regression analysis. The effectiveness of ethanol-based disinfectants at different concentrations was determined by in vitro and ex vivo evaluations. RESULTS: On plastic and skin surfaces, the Alpha, Beta, Delta, and Omicron variants exhibited approximately two-fold longer survival times than the Wuhan strain; the Omicron variants had the longest survival time. The median survival times of the Wuhan strain and the Alpha, Beta, Gamma, Delta, and Omicron (BA.1 and BA.2) variants on human skin surface were 8.6, 19.6, 19.1, 11.0, 16.8, 21.1, and 22.5 h, respectively. The in vitro evaluation showed that the Wuhan strain and the Alpha, Beta, Gamma, Delta, and Omicron (BA.1 and BA.2) variants were completely inactivated within 15 s by 32.5%, 35%, 35%, 32.5%, 35%, 40%, and 40% ethanol, respectively. However, all viruses on human skin were completely inactivated by exposure to 35% ethanol for 15 s. CONCLUSIONS: SARS-CoV-2 VOCs, especially the Omicron variants, have higher environmental stability than the Wuhan strain, increasing their transmission risk and contributing to their spread.

Alcohol abuse as a potential risk factor of solitary death among people living alone: a cross-sectional study in Kyoto, Japan.

BACKGROUND: Solitary death is an emerging public health problem in developed countries. Alcohol abuse is associated with social isolation and excess mortality. However, data on the association between alcohol abuse and solitary death are limited. Our purposes were to assess whether alcohol abuse is associated with a long interval from death to discovery among people living alone. METHODS: This is a cross-sectional study using the data on subjects from the largest forensic database in Kyoto, Japan, from February 2012 to December 2015. Solitary death was defined as a phenomenon of dying alone at home and remaining undiscovered for more than 1 week. All the subjects who lived alone and aged over 18 at the time of death were included in the study. The presence of alcohol abuse was identified via an investigation during home visits. Proportional ratios were calculated using a fitted logit model to evaluate the association of alcohol abuse on solitary death after adjusting for possible confounders. RESULTS: A total of 235 subjects were included in the analysis. The mean age (standard deviation) of subjects at the time of death was 63.4 (15.1) years, and approximately 61.8% and 38.9% of subjects in the alcohol and non-alcohol abuse groups, respectively, experienced solitary death. Multivariable analyses revealed that alcohol abuse was associated with solitary death (adjusted proportion ratio: 1.50; 95% confidence interval: 1.12-2.00). CONCLUSIONS: The findings of this study could help identify individuals at higher risk of solitary death. Moreover, calling the attention of people with alcohol abuse may be beneficial to prevent solitary death.

Higher Viral Stability and Ethanol Resistance of Avian Influenza A(H5N1) Virus on Human Skin.

Evaluating the stability of highly pathogenic avian influenza viruses on human skin and measuring the effectiveness of disinfectants are crucial for preventing contact disease transmission. We constructed an evaluation model using autopsy skin samples and evaluated factors that affect the stability and disinfectant effectiveness for various subtypes. The survival time of the avian influenza A(H5N1) virus on plastic surfaces was ≈26 hours and on skin surfaces ≈4.5 hours, >2.5-fold longer than other subtypes. The effectiveness of a relatively low ethanol concentration (32%-36% wt/wt) against the H5N1 subtype was substantially reduced compared with other subtypes. Moreover, recombinant viruses with the neuraminidase gene of H5N1 survived longer on plastic and skin surfaces than other recombinant viruses and were resistant to ethanol. Our results imply that the H5N1 subtype poses a higher contact transmission risk because of its higher stability and ethanol resistance, which might depend on the neuraminidase protein.

Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection.

Various chemical probes for the detection of reactive oxygen species have been developed to examine oxidative stress associated with different pathologies. L-012, a luminol-based chemiluminescent probe, is widely used to detect extracellular superoxide because of its high sensitivity. We herein demonstrated that the co-application of the peptide boronic acid proteasome inhibitor, bortezomib, with L-012 significantly increased its luminescence without affecting the background. More than a 5-fold increase was detected in the total luminescence of L-012 in both NADPH oxidase-expressing cells and the xanthine oxidase-dependent cell-free superoxide generation system, but not in their background. Therefore, bortezomib increased the signal-to-background ratio and improved the detection of low levels of superoxide. The application of MLN2238, another peptide boronic acid proteasome inhibitor, also enhanced the luminescence of L-012. In contrast, carfilzomib, an epoxyketone proteasome inhibitor, did not increase luminescence, suggesting that the effects of bortezomib depend on the chemical structure of the peptide boronic acid, but not on its pharmacological effects. Bortezomib-induced enhancements appeared to be specific to the detection of superoxide because the detection of H2O2 by Amplex Red/HRP was not affected by the application of bortezomib. In the quantitative detection of the superoxide-specific oxidative product 2-hydroxyethidium (2-OH-E+), the application of bortezomib resulted in a 2-fold increase in the level of 2-OH-E+. Therefore, bortezomib sensitizes the detection of superoxide in both cell-based and cell-free systems, highlighting a novel feature of compounds containing the peptide boronic acid as powerful enhancers for the detection of superoxide.

Evaluation of the Residual Disinfection Effects of Commonly Used Skin Disinfectants against Viruses: An Innovative Contact Transmission Control Method.

Lasting disinfection effects, that is, the residual disinfection effects (RDEs), of skin-coated disinfectants have rarely been considered for infection control owing to the challenges involved in the accurate evaluation of RDEs. In this study, we constructed a new skin evaluation model and determined the RDEs of existing disinfectants against viruses. Our results showed that ethanol and isopropanol had no RDE, whereas povidone-iodine, chlorhexidine gluconate, and benzalkonium chloride (BAC) exhibited RDEs, with 10% povidone-iodine and 0.2% BAC showing particularly strong RDEs. The RDE of 0.2% BAC was strong enough to reduce the median survival times of severe acute respiratory syndrome coronavirus-2, human coronavirus-OC43, and influenza virus from 670 to 5.2, 1300 to 12, and 120 to 4.2 min, respectively. Additionally, this strong RDE was maintained even 4 h after coating the skin. Clinical data also showed that the strong RDE of 0.2% BAC was maintained for more than 2 h. Thus, applying disinfectants with strong RDEs on the skin correlates with a reduction in virus survival time and appears to create a skin surface environment that is not conducive to virus survival. A prolonged reduction in virus survival decreases the contact transmission risk, thereby enabling stronger infection control.

Utilization of oral check-up data of autopsy cases.

The maintenance of oral health contributes to the overall health of an individual. Similar to the Adult Dental Health Survey in the UK, the Survey of Dental Diseases (SDD) is being conducted in Japan. However, our routine oral examination of autopsy cases showed that the oral cavities of the autopsied bodies were unhealthy as compared to the SDD data. Therefore, this study sought to understand the actual condition of the oral cavity and to clarify the factors related to the degradation of the oral status. This was achieved by comparing the data of the SDD subjects and autopsy cases in terms of the average number of teeth lost, the average total number of caries (number of decayed, missing, or filled teeth), percentage of people with untreated teeth, and 80-20 achievement rate (percentage of people aged 80 years with 20 or more teeth). Furthermore, this study investigated the relationship between the average number of missing teeth and smoking, psychiatric disorders, and body mass index (BMI) in autopsy cases. The oral condition of all autopsy cases was worse than that of SDD subjects. The 80-20 achievement rate was observed in more than half of the SDD subjects and only in less than 20% of the autopsy cases. Moreover, the average number of missing teeth was not significantly different depending on smoking status, BMI, and presence of psychiatric disorders in the autopsy cases. Unlike the SDD volunteers, there was no bias related to the inclusion of autopsy cases. Therefore, by analysing the data of both autopsy cases and SDD subjects, it may be possible to evaluate the oral condition more realistically. This could contribute to the realisation of improved health and quality of life.