The Effectiveness and Limitations of Triphenyltetrazolium Chloride to Detect Acute Myocardial Infarction at Forensic Autopsy

Kakimoto, Yu; Tsuruyama, Tatsuaki; Miyao, Masashi; Abiru, Hitoshi; Saito, Shinji; Kotani, Hirokazu; Haga, Hironori; Tamaki, Keiji

doi:10.1097/paf.0b013e31828879cd

Cited by 26 publications

(16 citation statements)

References 24 publications

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“…In fact, ischemic heart failure, the main cause of sudden cardiac death, scarcely displays any abnormal morphological changes 12 h after the initial acute myocardial infarction (AMI) [ 1 ]. Histological staining methods, such as triphenyltetrazolium chloride (TTC) staining, are often useful when visualizing the infarcted myocardial area during the sub-acute phase; however, these techniques are incapable of grossly detecting morphological changes in the early phase of AMI [ 2 ]. Therefore, it is essential to investigate novel biomarkers whose expression levels change rapidly during AMI.…”

Section: Introductionmentioning

confidence: 99%

MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients

et al. 2015

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MicroRNAs (miRNAs) are very short (18–24 nucleotides) nucleic acids that are expressed in a number of biological tissues and have been shown to be more resistant to extreme temperatures and pH compared to longer RNA molecules, like mRNAs. As miRNAs contribute to diverse biological process and respond to various kinds of cellular stress, their utility as diagnostic biomarkers and/or therapeutic targets has recently been explored. Here, we have evaluated the usefulness of miRNA quantification during postmortem examination of cardiac tissue from acute myocardial infarction (AMI) patients. Cardiac tissue was collected within one week of the patient’s death and either frozen (19 samples) or fixed in formalin for up to three years (36 samples). RNA integrity was evaluated with an electropherogram, and it appears that longer RNAs are fragmented after death in the long-term fixed samples. Quantitative PCR was also performed for seven miRNAs and three other small RNAs in order to determine the appropriate controls for our postmortem analysis. Our data indicate that miR-191 and miR-26b are more suitable than the other types of small RNA molecules as they are stably detected after death and long-term fixation. Further, we also applied our quantitation method, using these endogenous controls, to evaluate the expression of three previously identified miRNA biomarkers, miR-1, miR-208b, and miR-499a, in formalin-fixed tissues from AMI patients. Although miR-1 and miR-208b decreased (1.4-fold) and increased (1.2-fold), respectively, in the AMI samples compared to the controls, the significance of these changes was limited by our sample size. In contrast, the relative level of miR-499a was significantly decreased in the AMI samples (2.1-fold). This study highlights the stability of miRNAs after death and long-term fixation, validating their use as reliable biomarkers for AMI during postmortem examination.

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Section: Introductionmentioning

confidence: 99%

MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients

et al. 2015

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“…2,3,5-Triphenyltetrazolium chloride (TTC) is a redox indicator that produces water insoluble red formazan crystals when reduced [ 4 ]. It has been used extensively for several decades as an indicator of tissue ischaemia [ 5 , 6 ], and more recently to detect the presence of live micro-organisms based on metabolic activity of intracellular enzymes [ 4 ]. Accumulation of TTC within live bacteria provides high contrast between bacterial colonies and several solid agar media.…”

Section: Resultsmentioning

confidence: 99%

Validation of an automated colony counting system for group A Streptococcus

et al. 2016

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BackgroundThe practice of counting bacterial colony forming units on agar plates has long been used as a method to estimate the concentration of live bacteria in culture. However, due to the laborious and potentially error prone nature of this measurement technique, an alternative method is desirable. Recent technologic advancements have facilitated the development of automated colony counting systems, which reduce errors introduced during the manual counting process and recording of information. An additional benefit is the significant reduction in time taken to analyse colony counting data. Whilst automated counting procedures have been validated for a number of microorganisms, the process has not been successful for all bacteria due to the requirement for a relatively high contrast between bacterial colonies and growth medium. The purpose of this study was to validate an automated counting system for use with group A Streptococcus (GAS).ResultsMethods: Twenty-one different GAS strains, representative of major emm-types, were selected for assessment. In order to introduce the required contrast for automated counting, 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) dye was added to Todd–Hewitt broth with yeast extract (THY) agar. Growth on THY agar with TTC was compared with growth on blood agar and THY agar to ensure the dye was not detrimental to bacterial growth. Automated colony counts using a ProtoCOL 3 instrument were compared with manual counting to confirm accuracy over the stages of the growth cycle (latent, mid-log and stationary phases) and in a number of different assays. The average percentage differences between plating and counting methods were analysed using the Bland–Altman method.ConclusionsResults: A percentage difference of ±10 % was determined as the cut-off for a critical difference between plating and counting methods. All strains measured had an average difference of less than 10 % when plated on THY agar with TTC. This consistency was also observed over all phases of the growth cycle and when plated in blood following bactericidal assays. Agreement between these methods suggest the use of an automated colony counting technique for GAS will significantly reduce time spent counting bacteria to enable a more efficient and accurate measurement of bacteria concentration in culture.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-016-1875-z) contains supplementary material, which is available to authorized users.

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“…In an article concerning the effectiveness of triphenyltetrazolium chloride to detect acute myocardial infarction, the authors state that "it is not realistic to apply methods confirmed in animals" [68]. Stokes et al observed contrasting dynamics of mammalian analogues in experimental taphonomy, and state that "the results of this study showed many differences between porcine, bovine, and ovine SMTs (skeletal muscular tissues) when considering them as analogues for human SMT2 in decomposition soil studies" [69].…”

Section: Discussionmentioning

confidence: 96%

“…From this point of view, the history of forensic sciences has provided us with much evidence of the inapplicability of data obtained from studies performed on animal models, including the Daubert sentence [66] and several recent studies have again stressed the limits of this approach in different fields of forensic sciences, from the estimation of post mortem interval to genetics [67][68][69][70][71][72][73][74][75]. In an article concerning the effectiveness of triphenyltetrazolium chloride to detect acute myocardial infarction, the authors state that "it is not realistic to apply methods confirmed in animals" [68].…”

Section: Discussionmentioning

confidence: 99%