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Cx43 immunofluorescence. a (15 min after LAD ligation) Gap junctional expression in a small ischemic region. b (30 min after LAD ligation) Slightly more intense expression. A larger involved zone and a more intense fluorescence signal were observed, proportionally, at 1 h (c) and 2 h (d) after LAD ligation. No signal was detected in the non-ischemic zone (right ventricle, e)
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The post-mortem diagnosis of acute myocardial ischemia remains a challenge for both clinical and forensic pathologists. We performed an experimental study (ligation of left anterior descending coronary artery in rats) in order to identify early markers of myocardial ischemia, to further apply to forensic and clinical pathology in cases of sudden ca...
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The goal of this study was to assess whether early markers of myocardial ischemia, identified in a previous experimental work, can be applied in forensic pathology cases of sudden, ischemic cardiac death. These markers include desphosphorylated connexin 43 (Cx43), JunB, TUNEL assay, myoglobin, and troponin T. Fourteen cases of sudden cardiac death...
When exposed to oxidative and electrophilic stresses, a protective antioxidant response initiates through nuclear factor erythroid 2-related factor 2 (Nrf2). Nonetheless, 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, immunohis...
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, immunohistochemic...
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... Apart from proteins, metabolites and other small molecules, such as NEFAs and circRNAs, have demonstrated potential diagnostic value [14,15]. Furthermore, postmortem computed tomography coronary angiography has been utilized to identify the presence and severity of coronary artery disease and visualize areas of myocardial infarction [16][17][18]. In cases where autopsies yield negative findings, postmortem molecular autopsy, which investigates gene variants related to cardiac function or pathology (e.g., Long-QT syndrome, Brugada syndrome, and idiopathic ventricular fibrillation), currently represents the most effective method for explaining such sudden deaths [19]. ...
The aim of this study is to identify a rapid, sensitive, and non-destructive auxiliary approach for postmortem diagnosis of SCD, addressing the challenges faced in forensic practice.
ATR-FTIR spectroscopy was employed to collect spectral features of blood samples from different cases, combined with pathological changes. Mixed datasets were analyzed using ANN, KNN, RF, and SVM algorithms. Evaluation metrics such as accuracy, precision, recall, F1-score and confusion matrix were used to select the optimal algorithm and construct the postmortem diagnosis model for SCD.
A total of 77 cases were collected, including 43 cases in the SCD group and 34 cases in the non-SCD group. A total of 693 spectrogram were obtained. Compared to other algorithms, the SVM algorithm demonstrated the highest accuracy, reaching 95.83% based on spectral biomarkers. Furthermore, by combing spectral biomarkers with age, gender, and cardiac histopathological changes, the accuracy of the SVM model could get 100%.
Integrating artificial intelligence technology, pathology, and physical chemistry analysis of blood components can serve as an effective auxiliary method for postmortem diagnosis of SCD.
... Interpreting fibronectin (FN) levels can be challenging due to the influence of various factors such as resuscitation and overall hypoxia. On the other hand, while C5b-9 demonstrates high specificity and sensitivity, it may not account for all instances of suspected acute ischemic heart disease (AIHD) (8)(9)(10). Furthermore, FN and C5b-9 become positive 1 and 2 hours after the onset of ischemia, respectively (8,11,12). ...
... On the other hand, while C5b-9 demonstrates high specificity and sensitivity, it may not account for all instances of suspected acute ischemic heart disease (AIHD) (8)(9)(10). Furthermore, FN and C5b-9 become positive 1 and 2 hours after the onset of ischemia, respectively (8,11,12). Since accurately diagnosing acute ischemic heart disease (AIHD) within the initial 2-hour period remains a challenge, ongoing research endeavors are concentrated on enhancing the sensitivity and specificity of pathological AIHD diagnosis by investigating additional markers (13). ...
When exposed to oxidative and electrophilic stresses, a protective antioxidant response initiates through nuclear factor erythroid 2-related factor 2 (Nrf2). Nonetheless, 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. In this study, we investigated the immunohistochemical levels of Nrf2, FN, and C5b-9 in human cardiac samples to explore their forensic relevance in the identification of acute cardiac ischemia. The heart samples were obtained from 25 AIHD cases and 39 non-AIHD cases as controls. Nrf2 showed the localization 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. In addition, grading of positive portions for cardiac FN and C5b-9 in myocardium was also significantly enhanced in AIHD, compared to controls. Collectively, these results indicate that immunohistochemical investigation of Nrf2 combined with FN, and/or C5b-9 holds potential for identifying early-stage myocardial ischemic lesions in cases of sudden cardiac death.
... The loss of cTnT was non-uniform with greater loss at the periphery compared to the central regions of infarcted tissue [35]. Meta-analysis of pericardial fluid and serum cardiac troponin I, cTnI; or cardiac troponin T, cTnT; in the investigation of cardiac death and acute myocardial infarction, AMI (source: data extracted from [28] [31]; (D-I) [32]; (J-O) [33]). ...
The diagnosis of acute cardiac pathology is a clinical challenge in both the living and in the postmortem setting. Cardiac troponin (cTn) T and cardiac troponin I released from the contractile apparatus of cardiomyocytes into the circulation can be detected by sensitive and specific immunoassays and are the gold standard biochemical test for diagnosis of acute coronary syndromes (ACS). Recently with the advent of more sensitive detection methods, elevation in non-ACS has become apparent causing clinical confusion. In most cases, these elevations are related to subclinical cardiac damage and often confer poor prognosis in cTn-positive patients. Biomarkers of cardiomyocyte damage may be of value in routine hospital and medico-legal autopsy. A significant body of evidence has emerged since the late 1990s, assessing the clinical utility of cardiac troponin in biological fluids or in immunohistochemical staining of cardiac tissue to aid in the diagnosis of acute cardiac pathology when standard microscopic evidence is inconclusive. This chapter reviews the extensive literature on the subject and details the disparity between pericardial fluid and serum for the use of cTn in the postmortem setting.
... • Coagulation/fibrinolysis proteases may act as natural C3 and C5 convertases • Thrombin efficiently cleaved C5 at a newly identified, highly conserved R947 site, generating previously undescribed intermediates C5(T) and C5b(T) • C5b(T)-9 membrane attack complex has significantly more lytic activity than with C5b-9 • Complement activation in the presence of thrombin leads to assembly of C5bT • Lectin pathway inhibitors may be beneficial in patients with coagulation abnormalities/bleeding and incomplete response to C5 blockade • This can be extended to DIC mediated TMA as dysregulated coagulation cascade triggers secondary complement activation and some genetic mutations have been isolated in DIC linking to alternative pathway dysfunction and coagulation cascade mutations Cardiac/vascular emergencies [83][84][85][86] : ...
Over the last decade there have been numerous advances in both the diagnosis and treatment of transplant‐associated thrombotic microangiopathy (TA‐TMA). These are largely the result of an improved understanding of complement activation in TA‐TMA and the ability to prevent end organ injury and death with timely initiation of complement‐blocking therapies. In this article, we review our current understanding of the pathophysiology of TA‐TMA, particularly as it pertains to complement activation, endothelial injury, and clinical management. We then review novel complement‐blocking therapies that are currently under investigation for use in TA‐TMA, as well as discuss special considerations for complement‐blocking therapy in hematopoietic stem cell transplant recipients. Through these reviews we aim to answer or at least provide an educated discussion on the most commonly posed TA‐TMA management questions and dilemmas.
... In forensic practice, ischemic heart disease is a prevailing cause of SCDs, with atherosclerotic cardiovascular diseases (ASCVD) being the most recurrent substrate [18]. The triggering factors leading from ASCVD to the onset of SCD generally include transitory coronary spasms and prolonged occlusive coronary stenosis [19]. In cases of CAS-induced sudden deaths, several morphological changes within the coronary artery wall have been shown to reflect the occurrence of antemortem CAS. ...
... Pituitrin is extracted from the pituitary gland that contains vasopressin and oxytocin. Pituitrin is able to affect circulation and cause cardiac ischemia by inducing vascular contraction [19] and, therefore, has been widely used to create a CAS model [20,21]. We have previously established a CAS provocation model by the intraperitoneal (i.p.) injection of Pituitrin (5.1 U/kg) into mice with normal coronary arteries [25]. ...
Coronary artery spasm (CAS) plays an important role in the pathogenesis of many ischemic heart entities; however, there are no established diagnostic biomarkers for CAS in clinical and forensic settings. This present study aimed to identify such serum biomarkers by establishing a rabbit CAS provocation model and integrating quantitative serum proteomics, parallel reaction monitoring/mass spectrometry-based targeted proteomics, and partial least-squares discriminant analysis (PLS-DA). Our results suggested that SELENBP1 and VCL were potential candidate biomarkers for CAS. In independent clinical samples, SELENBP1 and VCL were validated to be significantly lower in serum but not blood cells from CAS patients, with the reasons for this possibly due to the decreased secretion from cardiomyocytes. The areas under the curve of the receiver operating characteristics (ROC) analysis were 0.9384 for SELENBP1 and 0.9180 for VCL when diagnosing CAS. The CAS risk decreased by 32.3% and 53.6% for every 10 unit increases in the serum SELENBP1 and VCL, respectively. In forensic samples, serum SELENBP1 alone diagnosed CAS-induced deaths at a sensitivity of 100.0% and specificity of 72.73%, and its combination with VCL yielded a diagnostic specificity of 100.0%, which was superior to the traditional biomarkers of cTnI and CK-MB. Therefore, serum SELENBP1 and VCL could be effective biomarkers for both the clinical and forensic diagnosis of CAS.
... At present, most studies on the determination of asphyxia use molecular biological techniques to find differentially expressed mRNAs, miRNAs or proteins, such as DUSP1/KCNJ2 (Zeng et al., 2018), miRNA-122 , miRNA-3185 (Han et al., 2020;Han et al., 2021), SP-A (Zhu et al., 2000), HIF-1a (Cecchi et al., 2014), GLUT1/VAGF (Zhao et al., 2008), AQP-5 , cytochrome C/AIF (Zhang et al., 2019) and MRP-8/-14 (Gutjahr and Madea, 2019). The same is true for SCD (Sabatasso et al., 2016, Campobasso et al., 2008, Carvajal-Zarrabal et al., 2017. However, these diagnostic biomarkers cannot be applied in practical work because their sensitivity and specificity need further investigation, as well as the effect of postmortem degradation. ...
It is important to determine the cause of death in the case of asphyxia. However, it is difficult to conclude death by asphyxia, especially when the deceased has underlying heart disease, because there are often no specific and representative corpse signs for both asphyxia and sudden cardiac death (SCD). The aim of the present work was to investigate the potential of metabolomics to discriminate asphyxia from SCD as the cause of death. A total of thirty male Sprague–Dawley rats were used to construct models of asphyxia, SCD (interfering cause of death), and cervical dislocation (control). Untargeted and widely targeted metabolomics approaches were used to obtain rat pulmonary metabolic profiles in this study. First, the metabolic alterations resulting from asphyxia were explored. There were significant changes found in carbohydrate metabolism, the endocrine system, and the sensory system. Second, we screened potential biomarkers and built classification models to determine the cause of death. Moreover, some biomarkers remained differentiated at 24 hours and 48 hours postmortem, so the cause of death could still be determined after death. This study showed the application potential of metabolomics to investigate the metabolic changes occurring in the process of death, as well as to determine the cause of death on the basis of metabolic differences even after death.
... Myocardial infarction, which is the most common cause of cardiovascular death worldwide, occurs with ischemic stress caused by coronary artery occlusion and cardiomyocyte death [1]. Pathological diagnosis of acute myocardial infarction can be difficult if death from ischemic injury has occurred within a short period of time [2]. Wavy fibers, hypereosinophilia, nuclear changes, which are among the earliest microscopic changes, may not be immediately noticeable. ...
... Recently, studies have been conducted to investigate the immunohistochemical methods that try to detect the molecules that accumulate in the cells or leak from the cells during ischemia with antibodies in order to show the damage to the cellular structures due to ischemia [2]. ...
... It has been shown that immunohistochemical methods are more suitable for forensic pathology practice in detecting early myocardial infarction [12]. Studies have also shown that this method supports the diagnosis of myocardial damage [2,8,12]. We also showed early myocardial infarction with immunohistochemical stains in our study. ...
Introduction: Pathological diagnosis of acute myocardial infarction can be difficult if death from ischemic injury has occurred within a short period of time. In this study, we aimed to determine the role of immunohistochemical markers in the diagnosis of early myocardial infarction.
Methods: The myocardium samples of 20 cases whose autopsies were performed at the Morgue Department of the Council of Forensic Medicine were evaluated. Hematoxylin and Eosin (H&E) stained slides and fibronectin, CD59, myoglobulin, troponin T, desmin, cathepsin S stained slides of 20 cases diagnosed with early myocardial infarction were retrospectively re-examined. The diagnosis of myocardial infarction was analyzed in two groups: Group 1: first eight hours, Group 2: 8-24 hours. The immunohistochemical staining patterns in these two groups were compared.
Results: Of the cases, 55% (n=11) had myocardial infarction consistent with the first eight hours, 45% (n=9) 8-24 hours with light microscopic examination. With fibronectin, 50% (n=10) of the cases showed Grade 1 staining, 5% (n=1) Grade 2, 15% (n=6) Grade 3 staining. The slides of three cases could not be reached. With CD59, 10% (n=2) of the cases showed Grade 1, 10% (n=2) Grade 2, 80% (n=16) Grade 3 staining. With troponin T, 50% (n=10) of the cases showed Grade 1, 45% (n=9) Grade 2, 5% (n=1) Grade 3 depletion. With cathepsin S, 10% (n=2) of the cases showed Grade 1 and 80% (n=16) Grade 3 depletion. The slides of two cases could not be reached. With desmin, 75% (n=15) had Grade 1 and 25% (n=5) Grade 2 depletion. Grade 3 depletion with myoglobulin was observed in all cases.
Conclusion: The diagnosis of early myocardial infarction, which may pose a problem for the forensic pathologist, may become easier with immunohistochemical methods. In cases where morphological findings are insufficient, it is more useful for diagnosis to be applied as a panel.
... Holmbom et al. found close correspondence of antifibronectin staining with the infarct size from tetrazolium-stained slices in a pig MI model [40]. More recently, fibronectin expression was identified as one of the early markers for identifying MI as a cause of death in human post mortem samples [41]. In MI patients, fibronectin appears in the ischemic cardiomyocytes within a day and disappears gradually as the scar tissue develops [42]. ...
Location and extent of necrosis are valuable information in the management of myocardial infarction (MI). Methods. We investigated 2-deoxy-2-¹⁸F-fluoro glucaric acid (FGA), a novel infarct-avid agent, for positron emission tomography (PET) of MI. We synthesized FGA from commercially available ¹⁸F-fluoro-2-deoxy-2-D-glucose (FDG). MI was induced in mice by permanently occluding the left anterior descending coronary artery. Biodistribution of FGA was assessed 1 h after FGA injection (11 MBq). PET/CT was conducted 1 h, 6 h, 1 d, 3 d, and 4 d after MI. Subcellular compartment of FGA accumulation in necrosis was studied by tracing the uptake of biotin-labeled glucaric acid with streptavidin-HRP in H2O2-treated H9c2 cardiomyoblasts. Streptavidin-reactive protein bands were identified by LC-MS/MS. Results. We obtained a quantitative yield of FGA from FDG within 7 min (radiochemical purity>99%). Cardiac uptake of FGA was significantly higher in MI mice than that in control mice. Imaging after 1 h of FGA injection delineated MI for 3 days after MI induction, with negligible background signal from surrounding tissues. Myocardial injury was verified by tetrazolium staining and plasma troponin (47.63 pg/mL control versus 311.77 pg/mL MI). In necrotic H9c2 myoblasts, biotinylated glucaric acid accumulated in nuclear fraction. LC-MS/MS primarily identified fibronectin in necrotic cells as a putative high fidelity target of glucaric acid. Conclusion. FGA/PET detects infarct early after onset of MI and FGA accumulation in infarct persists for 3 days. Its retention in necrotic cells appears to be a result of interaction with fibronectin that is known to accumulate in injured cardiac tissue.
... The concentration of Troponin-I, LDH and CPK in the blood is increased following myocardial damage, such as acute myocardial infarction, shock, hypoxia or acute coronary artery disease. Therefore, the aforementioned factors can be used as the clinical diagnosis of myocardial infarction and evaluation (45). ...
Swimming is important for promoting and maintaining health, as it can increase the efficiency of the cardiovascular system and decrease the occurrence of cardiovascular diseases. The objective of the present study was to examine whether swimming training could decrease myocardial injury in rats caused by myocardial ischemia/reperfusion (I/R). Sprague-Dawley rats were randomized into four groups, namely the Sham, coronary artery occlusion, swimming training and ischemic preconditioning (IPC) groups. Myocardial I/R was induced in anesthetized male Sprague-Dawley rats by a 40-min occlusion followed by a 3-h reperfusion of the left anterior descending coronary artery. The rats were sacrificed after surgery and their hearts were examined. The results demonstrated that the number of TUNEL-positive nuclei and degree of caspase-3 activation were both significantly increased in the myocardium following myocardial I/R in rats, indicating increased cardiomyocyte apoptosis. On the other hand, swimming training decreased the serum levels of creatine phosphokinase, lactate dehydrogenase and cardiac troponin I, and was associated with reduced histological damage and myocardial infarct size. Furthermore, swimming training also reduced TNF-α levels, caspase-3 activation and enhanced Bcl-2 activation, which decreased the number of apoptotic cells in the myocardium. The findings of the present study showed that swimming training and IPC could similarly decrease myocardial injury following myocardial I/R, and may therefore be used as exercise training to effectively prevent myocardial injury.
... Myocardial infarction (MI), pathologically defined as myocardial cell death due to prolonged ischemia, is most representative acute ischemic heart disease (AIHD). After the onset of myocardial ischemia, histological cell death does not occur immediately, and it takes a finite period of time to develop-as little as 20 min, or less in some animal models [3][4][5] . Depending on the sensitivity of cardiomyocytes, macroscopic or microscopic postmortem examination takes several hours to identify myocardial necrosis. ...
... Recent works have immunohistochemically investigated, some markers such as fibronectin, C5b-9 and myoglobin which accumulate in or leak from cardiomyocytes after ischemia. However, these molecules almost failed to be detected in the very early phase of myocardial ischemia 5,[15][16][17][18] . ...
... C5b-9 is most commonly used to support the final diagnosis of SCD because it can reveal areas of myocardial necrosis 5,15,27,30 . This property of C5b-9 is due to its direct involvement in the complement cascade. ...
Heme oxygenase-1 (HO-1), an inducible stress-response protein, exerts anti-oxidant and anti-apoptotic effects. However, its significance in forensic diagnosis of acute ischemic heart diseases (AIHD) such as myocardial infarction (MI) is still unknown. We examined the immunohistochemical expression of HO-1 in the heart samples to discuss their forensic significance to determine acute cardiac ischemia. The heart samples were obtained from 23 AIHD cases and 33 non-AIHD cases as controls. HO-1 positive signals in cardiomyocyte nuclear were detected in 78.2% of AIHD cases, however, that were detected in only 24.2% control cases with statistical difference between AIHD and non-AIHD groups. In contrast to HO-1 protein expression, there was no significant difference in the appearance of myoglobin pallor regions and leukocyte infiltration in the hearts between AIHD and non-AIHD groups. From the viewpoints of forensic pathology, intracardiac HO-1 expression would be considered a valuable marker to diagnose AIHD as the cause of death.
