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Article: Unexpected sudden death on arrival in a healthy middle-aged man associated with COVID-19-related diffuse cardiac injury: A case report.

Isozaki, Shotaro / Kakimoto, Yu / Ikeda, Haruka / Matsushima, Yutaka / Tsuboi, Akio / Osawa, Motoki

2023 Volume 10, Issue 1, Page(s) e23460

Abstract: Coronavirus disease 2019 (COVID-19) is an emerging respiratory infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 has been reported to lead to acute cardiac injury, but previous research indicated that the ...

Abstract	Coronavirus disease 2019 (COVID-19) is an emerging respiratory infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 has been reported to lead to acute cardiac injury, but previous research indicated that the mechanism is different from that of other viruses and remains poorly understood. Herein, we describe a case of COVID-19-associated sudden death, in a healthy 47-year-old man after developing diffuse cardiac necrosis. Two days before death, the patient developed general malaise without respiratory symptoms. The patient's fatigue worsened with time, and he ultimately developed cardiac arrest in an ambulance; however, resuscitation was unsuccessful. Antigen testing performed at the hospital revealed that the patient was positive for SARS-CoV-2 virus. At autopsy, contraction band necrosis was observed insularly in all areas of the myocardium. CD42b immunohistochemical staining indicated platelet aggregation in the microvessels around the cardiac necrosis area, suggesting COVID-19 can be fatal for healthy people by microcirculatory disturbance due to diffuse cardiac injury arising from platelet activiation. This unique mechanism can be a novel therapeutic target of COVID-19-related cardiac injury.
Language	English
Publishing date	2023-12-10
Publishing country	England
Document type	Case Reports
ZDB-ID	2835763-2
ISSN	2405-8440
ISSN	2405-8440
DOI	10.1016/j.heliyon.2023.e23460
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Article ; Online: A case of a motor vehicle collision suspected as associated with development of angiosarcoma.

Ikeda, Haruka / Isozaki, Shotaro / Kakimoto, Yu / Ueda, Atsushi / Tsuboi, Akio / Osawa, Motoki

Legal medicine (Tokyo, Japan)

2023 Volume 67, Page(s) 102332

Abstract: Trauma has been suspected as a factor leading to development of angiosarcoma, a malignant soft-tissue tumor. We conducted a forensic autopsy to investigate a putative relation between a motor vehicle collision and the driver's later death from ... ...

Abstract	Trauma has been suspected as a factor leading to development of angiosarcoma, a malignant soft-tissue tumor. We conducted a forensic autopsy to investigate a putative relation between a motor vehicle collision and the driver's later death from angiosarcoma. A vehicle operated by a man in his 60 s collided with an oncoming vehicle at a curve. The victim noticed no injury at the scene. However, 45 days later, he was transferred to an emergency room with dyspnea and bloody sputum. After diagnosis of angiosarcoma, he died of respiratory failure 132 days later. The bereaved family speculated about a relation between the collision and angiosarcoma onset. At autopsy, tumor cells of the scalp had metastasized to the lung, pleura, liver, and spleen. Histopathological examinations revealed characteristic features of angiosarcoma with positive immune-staining for CD31, CD34, and factor VIII. When a person dies some time after a collision, it is designated as a delayed death. In such cases, the relevance of trauma to the person's death is often an issue of concern. Because the interval between trauma and angiosarcoma development was short, only 45 days, the angiosarcoma might be coincidental. Therefore, we rejected the relation. Forensic experts sometimes need to investigate such inquiries.
MeSH term(s)	Male ; Humans ; Hemangiosarcoma/diagnosis ; Lung/pathology ; Motor Vehicles
Language	English
Publishing date	2023-10-09
Publishing country	Ireland
Document type	Case Reports
ZDB-ID	2019555-2
ISSN	1873-4162 ; 1344-6223
ISSN (online)	1873-4162
ISSN	1344-6223
DOI	10.1016/j.legalmed.2023.102332
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Article ; Online: Proteomic profiling of sudden cardiac death with acquired cardiac hypertrophy.

Kakimoto, Yu / Ueda, Atsushi / Ito, Masatoshi / Tanaka, Masayuki / Kubota, Tomoko / Isozaki, Shotaro / Osawa, Motoki

International journal of legal medicine

2023 Volume 137, Issue 5, Page(s) 1453–1461

Abstract: Background: Cardiac hypertrophy, which develops in middle-aged and older individuals as a consequence of hypertension and obesity, is an established risk factor for sudden cardiac death (SCD). However, it is sometimes difficult to differentiate SCD with ...

Abstract	Background: Cardiac hypertrophy, which develops in middle-aged and older individuals as a consequence of hypertension and obesity, is an established risk factor for sudden cardiac death (SCD). However, it is sometimes difficult to differentiate SCD with acquired cardiac hypertrophy (SCH) from compensated cardiac hypertrophy (CCH), at autopsy. We aimed to elucidate the proteomic alteration in SCH, which can be a guideline for future postmortem diagnosis. Methods: Cardiac tissues were sampled at autopsy. SCH group consisted of ischemic heart failure, hypertensive heart failure, and aortic stenosis. CCH group included cases of non-cardiac death with cardiac hypertrophy. The control group comprised cases of non-cardiac death without cardiac hypertrophy. All patients were aged > 40 years, and hypertrophic cardiomyopathy was not included in this study. We performed histological examination and shotgun proteomic analysis, followed by quantitative polymerase chain reaction analysis. Results: Significant obesity and myocardial hypertrophy, and mild myocardial fibrosis were comparable in SCH and CCH cases compared to control cases. The proteomic profile of SCH cases was distinguishable from those of CCH and control cases, and many sarcomere proteins were increased in SCH cases. Especially, the protein and mRNA levels of MYH7 and MYL3 were significantly increased in SCH cases. Conclusion: This is the first report of cardiac proteomic analysis in SCH and CCH cases. The stepwise upregulation of sarcomere proteins may increase the risk for SCD in acquired cardiac hypertrophy before cardiac fibrosis progresses significantly. These findings can possibly aid in the postmortem diagnosis of SCH in middle-aged and older individuals.
MeSH term(s)	Middle Aged ; Humans ; Aged ; Proteomics ; Death, Sudden, Cardiac/etiology ; Death, Sudden, Cardiac/pathology ; Cardiomyopathies ; Heart Failure ; Fibrosis ; Hypertension/complications ; Obesity ; Cardiomegaly
Language	English
Publishing date	2023-06-07
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1055109-8
ISSN	1437-1596 ; 0937-9827
ISSN (online)	1437-1596
ISSN	0937-9827
DOI	10.1007/s00414-023-03038-6
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Article: Soluble thrombomodulin ameliorates aberrant hemostasis after rewarming in a rat accidental hypothermia model

Takauji, Shuhei / Tanaka, Hiroki / Hayakawa, Mineji / Horioka, Kie / Isozaki, Shotaro / Konishi, Hiroaki

Biochemical and biophysical research communications. 2022 Jan. 08, v. 587

2022

Abstract: Accidental hypothermia (AH) sometimes leads to coagulation disorder, especially in severe AH. We previously demonstrated that intrasplenic platelet activation caused aberrant hemostasis and thrombus formation after rewarming in a murine AH model. However, ...

Abstract	Accidental hypothermia (AH) sometimes leads to coagulation disorder, especially in severe AH. We previously demonstrated that intrasplenic platelet activation caused aberrant hemostasis and thrombus formation after rewarming in a murine AH model. However, no study has focused on the appropriate management of platelets causing coagulation activation after rewarming of AH. We investigated whether or not recombinant soluble thrombomodulin (rTM) can suppress thrombosis formation after rewarming using a rat AH model. Wistar rats were exposed to an ambient temperature of −20 °C under general anesthesia until their rectal temperature decreased to 26 °C. The Hypo group rats (n = 5) were immediately euthanized, while the Hypo/Re group (n = 5) and rTM group rats (n = 5), which were administered rTM (1 mg/kg) via the tail vein, were rewarmed until the rectal temperature returned to 34 °C and then euthanized 6 h later. Tissue and blood samples were collected from all rats for histopathological and coagulation analyses at euthanasia. There was no significant change in the D-dimer level in the Hypo group rats, while the D-dimer level was significantly elevated at 6 h after rewarming in the Hypo/Re group rats (P = 0.015), and histopathology detected both fibrin and platelets in the renal glomerulus. However, the rTM group rats did not show any elevation of the D-dimer levels at 6 h after rewarming, and no fibrin was noted on histopathology. rTM may be useful as an appropriate anticoagulant in cases of aberrant hemostasis after rewarming of AH.
Keywords	ambient temperature ; anesthesia ; anticoagulants ; caudal vein ; coagulation ; fibrin ; histopathology ; hypothermia ; mice ; models ; platelet activation ; rats ; research ; thrombosis
Language	English
Dates of publication	2022-0108
Size	p. 1-8.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	205723-2
ISSN	0006-291X ; 0006-291X
ISSN (online)	0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2021.11.086
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Article ; Online: Probiotic-derived heptelidic acid exerts antitumor effects on extraintestinal melanoma through glyceraldehyde-3-phosphate dehydrogenase activity control.

Isozaki, Shotaro / Konishi, Hiroaki / Tanaka, Hiroki / Yamamura, Chikage / Moriichi, Kentaro / Ogawa, Naoki / Fujiya, Mikihiro

BMC microbiology

2022 Volume 22, Issue 1, Page(s) 110

Abstract: Background: Several microorganisms inhabit the mammalian gastrointestinal tract and are associated with the pathogenesis of various diseases, including cancer. Recent studies have indicated that several probiotics produce antitumor molecules and inhibit ...

Abstract	Background: Several microorganisms inhabit the mammalian gastrointestinal tract and are associated with the pathogenesis of various diseases, including cancer. Recent studies have indicated that several probiotics produce antitumor molecules and inhibit host tumor progression. We demonstrated that heptelidic acid (HA), a sesquiterpene lactone derived from the probiotic Aspergillus oryzae, exerts antitumor effects against pancreatic cancer in vitro and in vivo. In this study, the antitumor effects of HA against extraintestinal melanoma were assessed in vitro and in vivo. Results: Sulforhodamine B (SRB) assay revealed that the growth of B16F10 cells was significantly inhibited by HA in a concentration-dependent manner. The enzymatic activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) decreased in proportion with the growth inhibition effect of HA. Moreover, oral HA administration significantly suppressed the growth of transplanted B16F10 tumors without any significant changes in biochemical test values. Moreover, GAPDH activity in the transplanted tumor tissues in the HA group significantly decreased compared with that in the PBS group. Conclusion: This study suggests that orally administered HA was absorbed in the gastrointestinal tract, reached the cancer cells transplanted in the skin, and inhibited GAPDH activity, thereby inhibiting the growth of extraintestinal melanoma cells. Thus, this study proposes a novel system for extraintestinal tumor regulation via gut bacteria-derived bioactive mediators.
MeSH term(s)	Animals ; Glyceraldehyde-3-Phosphate Dehydrogenases/chemistry ; Mammals ; Melanoma/drug therapy ; Probiotics/pharmacology ; Sesquiterpenes
Chemical Substances	Sesquiterpenes ; heptelidic acid (57710-57-3) ; Glyceraldehyde-3-Phosphate Dehydrogenases (EC 1.2.1.-)
Language	English
Publishing date	2022-04-22
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2041505-9
ISSN	1471-2180 ; 1471-2180
ISSN (online)	1471-2180
ISSN	1471-2180
DOI	10.1186/s12866-022-02530-0
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Article ; Online: An instance of homicide by electrocution with hand-made electrode plates.

Kakimoto, Yu / Ikeda, Haruka / Matsushima, Yutaka / Tsuboi, Akio / Ueda, Atsushi / Isozaki, Shotaro / Osawa, Motoki

Forensic science, medicine, and pathology

2022 Volume 19, Issue 4, Page(s) 568–572

Abstract: Homicide by electrocution is rare in forensics, and the identification of the cause of death can be quite difficult when the electric device is removed from the scene. We present an instance where the police were unsure of homicide in the initial ... ...

Abstract	Homicide by electrocution is rare in forensics, and the identification of the cause of death can be quite difficult when the electric device is removed from the scene. We present an instance where the police were unsure of homicide in the initial investigation. The offender used hand-made electrode plates for electrocution, which produced unique electric marks different from those produced by common electric devices such as electric wires. To the best of our knowledge, this is the first report of homicide by electrocution with electrode plates. We believe that the macroscopic and microscopic findings in this instance are quite valuable for forensic practitioners.
MeSH term(s)	Humans ; Electric Injuries ; Homicide ; Forensic Medicine ; Forensic Pathology
Language	English
Publishing date	2022-12-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	2195904-3
ISSN	1556-2891 ; 1547-769X
ISSN (online)	1556-2891
ISSN	1547-769X
DOI	10.1007/s12024-022-00566-2
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Article ; Online: DNA methylation-based age estimation and quantification of the degradation levels of bisulfite-converted DNA.

Shiga, Mihiro / Asari, Masaru / Takahashi, Yuta / Isozaki, Shotaro / Hoshina, Chisato / Mori, Kanae / Namba, Ryo / Okuda, Katsuhiro / Shimizu, Keiko

Legal medicine (Tokyo, Japan)

2023 Volume 67, Page(s) 102336

Abstract: DNA methylation modifications are known to influence epigenetic phenomena and have been a focus of forensic science research for some time. Degraded DNA after bisulfite treatment is widely used in DNA methylation analysis. In this study, we analyzed ... ...

Abstract	DNA methylation modifications are known to influence epigenetic phenomena and have been a focus of forensic science research for some time. Degraded DNA after bisulfite treatment is widely used in DNA methylation analysis. In this study, we analyzed methylation levels at 12 CpG sites of four selected genomic regions by pyrosequencing after bisulfite treatment. DNA was extracted from buccal swab samples collected from 102 Japanese individuals who were 21-77 years old. We also developed a simple method to quantify the degradation levels of bisulfite-converted DNA by real-time PCR, and evaluated the effect of DNA degradation on age estimation. We found that the methylation levels and chronological ages were highly correlated in the four selected regions, and the mean absolute deviation (MAD) between chronological and estimated ages was low at 3.88 years. These results indicated that pyrosequencing analysis at the 12 CpGs was useful for age estimation in the Japanese population. To develop a sensitive quantification method, we analyzed the amplification efficiency of short and long fragments from 10 regions by real-time PCR. The amplification efficiency was highest for CCDC102B, and the degradation levels of bisulfite-converted DNA for the 102 samples were categorized as moderately or heavily degraded. For the younger age groups (20-49 years), the MADs were lower for moderately degraded DNA than they were for heavily degraded DNA. This finding indicates that degradation levels affected the accuracy of age estimation in most of the samples; the exception was the samples from the 50-77 years age group.
MeSH term(s)	Humans ; Young Adult ; Adult ; Middle Aged ; Aged ; DNA Methylation/genetics ; Aging/genetics ; CpG Islands/genetics ; DNA/genetics ; Forensic Genetics/methods ; Sulfites
Chemical Substances	hydrogen sulfite (OJ9787WBLU) ; DNA (9007-49-2) ; Sulfites
Language	English
Publishing date	2023-10-20
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	2019555-2
ISSN	1873-4162 ; 1344-6223
ISSN (online)	1873-4162
ISSN	1344-6223
DOI	10.1016/j.legalmed.2023.102336
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Article ; Online: Soluble thrombomodulin ameliorates aberrant hemostasis after rewarming in a rat accidental hypothermia model.

Takauji, Shuhei / Tanaka, Hiroki / Hayakawa, Mineji / Horioka, Kie / Isozaki, Shotaro / Konishi, Hiroaki

Biochemical and biophysical research communications

2021 Volume 587, Page(s) 1–8

Abstract: Background: Accidental hypothermia (AH) sometimes leads to coagulation disorder, especially in severe AH. We previously demonstrated that intrasplenic platelet activation caused aberrant hemostasis and thrombus formation after rewarming in a murine AH ... ...

Abstract	Background: Accidental hypothermia (AH) sometimes leads to coagulation disorder, especially in severe AH. We previously demonstrated that intrasplenic platelet activation caused aberrant hemostasis and thrombus formation after rewarming in a murine AH model. However, no study has focused on the appropriate management of platelets causing coagulation activation after rewarming of AH. We investigated whether or not recombinant soluble thrombomodulin (rTM) can suppress thrombosis formation after rewarming using a rat AH model. Methods: Wistar rats were exposed to an ambient temperature of -20 °C under general anesthesia until their rectal temperature decreased to 26 °C. The Hypo group rats (n = 5) were immediately euthanized, while the Hypo/Re group (n = 5) and rTM group rats (n = 5), which were administered rTM (1 mg/kg) via the tail vein, were rewarmed until the rectal temperature returned to 34 °C and then euthanized 6 h later. Tissue and blood samples were collected from all rats for histopathological and coagulation analyses at euthanasia. Results: There was no significant change in the D-dimer level in the Hypo group rats, while the D-dimer level was significantly elevated at 6 h after rewarming in the Hypo/Re group rats (P = 0.015), and histopathology detected both fibrin and platelets in the renal glomerulus. However, the rTM group rats did not show any elevation of the D-dimer levels at 6 h after rewarming, and no fibrin was noted on histopathology. Conclusions: rTM may be useful as an appropriate anticoagulant in cases of aberrant hemostasis after rewarming of AH.
MeSH term(s)	Animals ; Anticoagulants/pharmacology ; Biomarkers/metabolism ; Blood Platelets/drug effects ; Blood Platelets/metabolism ; Blood Platelets/pathology ; Disease Models, Animal ; Fibrin/chemistry ; Fibrin/metabolism ; Fibrin Fibrinogen Degradation Products/metabolism ; Hypothermia/blood ; Hypothermia/complications ; Hypothermia/physiopathology ; Kidney Glomerulus/blood supply ; Kidney Glomerulus/drug effects ; Kidney Glomerulus/metabolism ; Kidney Glomerulus/pathology ; Male ; Platelet Activation/drug effects ; Rats ; Rats, Wistar ; Recombinant Proteins/pharmacology ; Rewarming/adverse effects ; Solubility ; Spleen/blood supply ; Spleen/drug effects ; Spleen/metabolism ; Spleen/pathology ; Thrombomodulin/administration & dosage ; Thrombosis/blood ; Thrombosis/etiology ; Thrombosis/physiopathology ; Thrombosis/prevention & control
Chemical Substances	Anticoagulants ; Biomarkers ; Fibrin Fibrinogen Degradation Products ; Recombinant Proteins ; Thrombomodulin ; fibrin fragment D ; Fibrin (9001-31-4)
Language	English
Publishing date	2021-11-26
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	205723-2
ISSN	1090-2104 ; 0006-291X ; 0006-291X
ISSN (online)	1090-2104 ; 0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2021.11.086
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Article ; Online: Age prediction by methylation analysis of small amounts of DNA using locked nucleic acids.

Takahashi, Yuta / Asari, Masaru / Isozaki, Shotaro / Hoshina, Chisato / Okuda, Katsuhiro / Mori, Kanae / Namba, Ryo / Ochiai, Wataru / Shimizu, Keiko

Journal of forensic sciences

2022

Abstract: Age prediction based on methylation analysis has been reported in many populations, with 10 ng or more of DNA usually required for each determination. In this study, we designed thermostable locked nucleic acid (LNA) primers by replacing a small number ... ...

Abstract	Age prediction based on methylation analysis has been reported in many populations, with 10 ng or more of DNA usually required for each determination. In this study, we designed thermostable locked nucleic acid (LNA) primers by replacing a small number of DNA bases in standard DNA primers with LNAs. We evaluated these primer sets by single-base extension analysis using 10, 5, or 2 ng of DNA that would be less than template DNA used in standard methylation testing, and determined sensitivity and accuracy. We analyzed EDARADD, SST, and KLF14 genes, targeting one CpG site in each gene. Melting temperature values of most LNA primers were 4°C higher than those of DNA primers. The intensities of signals from the EDARADD and SST genes were significantly improved by the LNA primers, by 3.3 times and 1.4 times, respectively, compared with the DNA primers using 2 ng of DNA. Coefficient of variation (CV) analysis was used to assess the accuracy of the determined methylation levels. CVs were increased using small amounts of DNA, but lower CVs were detected using LNA primers. We also showed high accuracy of age prediction for 51 individuals using LNA primers. The lowest mean absolute deviation was obtained using 10 ng of DNA and was 3.88 years with the LNA primers. Thermostable PCR primers were simply designed, and the LNAs improved the sensitivity and accuracy of methylation analysis for 10 ng or less of DNA.
Language	English
Publishing date	2022-09-23
Publishing country	United States
Document type	Journal Article
ZDB-ID	219216-0
ISSN	1556-4029 ; 0022-1198
ISSN (online)	1556-4029
ISSN	0022-1198
DOI	10.1111/1556-4029.15144
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Article ; Online: Hypoxia-induced nuclear translocation of β-catenin in the healing process of frostbite.

Isozaki, Shotaro / Tanaka, Hiroki / Horioka, Kie / Konishi, Hiroaki / Kashima, Shin / Takauji, Shuhei / Fujiya, Mikihiro / Druid, Henrik

Biochimica et biophysica acta. Molecular basis of disease

2022 Volume 1868, Issue 6, Page(s) 166385

Abstract: Frostbite occurs when the skin is exposed to localized low temperatures. The main causes of frostbite are thought to be direct cell injury due to freezing of cells and tissue ischemia due to abnormal blood circulation. However, the molecular mechanism of ...

Abstract	Frostbite occurs when the skin is exposed to localized low temperatures. The main causes of frostbite are thought to be direct cell injury due to freezing of cells and tissue ischemia due to abnormal blood circulation. However, the molecular mechanism of frostbite has not been elucidated. This study aims to explain the molecular dynamics of frostbite using a mouse frostbite model and keratinocyte cell culture. Comprehensive gene expression analysis performed on mouse skin samples revealed that β-catenin signaling is activated by frostbite. Immunohistochemistry showed nuclear translocation of β-catenin in the skin of frostbite model mice that was not observed in mice subjected to a mechanical skin damage model induced by tape stripping. Tissue hypoxia, as detected by pimonidazole staining, coexisted with nuclear expression of β-catenin. In keratinocyte cell cultures, nuclear translocation of β-catenin was induced by hypoxia, but not by low temperature. Hypoxia induced epithelial-mesenchymal transition - an important biological event in the healing process of skin - and in vitro wound-healing activity, both of which were suppressed by β-catenin inhibition. Our results suggest that during frostbite, impaired blood flow causes hypoxia, which in turn activates β-catenin that promotes keratinocyte motility and tissue repair.
MeSH term(s)	Frostbite/metabolism ; Humans ; Hypoxia/metabolism ; Keratinocytes/metabolism ; Wound Healing/physiology ; beta Catenin/metabolism
Chemical Substances	beta Catenin
Language	English
Publishing date	2022-03-12
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	60-7
ISSN	1879-260X ; 1879-2596 ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1879-260X ; 1879-2596 ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/j.bbadis.2022.166385
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