Degradation of Kidney and Psoas Muscle Proteins as Indicators of Post-Mortem Interval in a Rat Model, with Use of Lateral Flow Technology

Lee, Dong-Gi; Yang, Kyeong Eun; Hwang, Jeong Won; Kang, Ha‐Young; Lee, Seung Yeul; Choi, Seoyeon; Shin, Jong‐Shik; Jang, Ik Soon; An, Hyun Joo; Chung, Heesun; Jung, Hyo Il; Choi, Jong Soon

doi:10.1371/journal.pone.0160557

Cited by 28 publications

(23 citation statements)

References 39 publications

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“…However, from the low number of human samples in this study, it cannot be concluded whether there is a statistical decrease of the native band in later postmortem stages, as it was the case in rat vastus lateralis muscle. The significant decomposition of GAPDH protein in rats over a PMI of 96 hpm is confirmed by the proteomic analysis and by a study reporting postmortem GAPDH decrease in rat psoas muscle [ 17 ] over a similar period of time. Despite the trend to decrease, the presence of the native band of GAPDH band at a PMI of 96 hpm and the occurring degradation products at a larger PMI in the human sample H3 indicate possible applicability as a marker in later postmortem stages.…”

Section: Discussionmentioning

confidence: 60%

“…Postmortem changes, such as the loss of the native protein band or the occurrence of specific degradation products, were additionally shown to significantly correlate with the PMI. Among these proteins are calcineurin A, protein phosphatase 2A [ 13 ], titin, nebulin [ 14 ], desmin, troponin [ 15 ], vinculin [ 16 ], AMP-activated protein kinase, caspase 3, and glycogen synthase [ 17 ]. Furthermore, PMI-dependent desmin and cardiac troponin T degradation has been confirmed in humans [ 15 ] and was already successfully applied to obtain evidence about the succession of events in a criminal case [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

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Postmortem proteomics to discover biomarkers for forensic PMI estimation

et al. 2019

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The assessment of postmortem degradation of skeletal muscle proteins has emerged as a novel approach to estimate the time since death in the early to mid-postmortem phase (approximately 24 h postmortem (hpm) to 120 hpm). Current protein-based methods are limited to a small number of skeletal muscle proteins, shown to undergo proteolysis after death. In this study, we investigated the usability of a target-based and unbiased system-wide protein analysis to gain further insights into systemic postmortem protein alterations and to identify additional markers for postmortem interval (PMI) delimitation. We performed proteomic profiling to globally analyze postmortem alterations of the rat and mouse skeletal muscle proteome at defined time points (0, 24, 48, 72, and 96 hpm), harnessing a mass spectrometry-based quantitative proteomics approach. Hierarchical clustering analysis for a total of 579 (rat) and 896 (mouse) quantified proteins revealed differentially expressed proteins during the investigated postmortem period. We further focused on two selected proteins (eEF1A2 and GAPDH), which were shown to consistently degrade postmortem in both rat and mouse, suggesting conserved intra- and interspecies degradation behavior, and thus preserved association with the PMI and possible transferability to humans. In turn, we validated the usefulness of these new markers by classical Western blot experiments in a rat model and in human autopsy cases. Our results demonstrate the feasibility of mass spectrometry–based analysis to discover novel protein markers for PMI estimation and show that the proteins eEF1A2 and GAPDH appear to be valuable markers for PMI estimation in humans. Electronic supplementary material The online version of this article (10.1007/s00414-019-02011-6) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 60%

Section: Introductionmentioning

confidence: 99%

Postmortem proteomics to discover biomarkers for forensic PMI estimation

et al. 2019

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“…This constitutes a major advantage in both research (e.g., multiple samples can be taken and analyzed, without interference by previous samplings) and practical application (e.g., even in severely injured bodies or body parts, there is a high chance to be able to collect unaffected tissue for analysis). Available data, however, is largely limited to thigh muscle [ 5 – 8 ], with few exceptions (e.g., M. psoas [ 9 ], M. gastrocnemius [ 10 ]). To fully benefit from the described advantages of muscle tissue, further research is required regarding decomposition similarities and deviations within individual muscles, between muscles, and between muscle types (skeletal muscle, cardiac and smooth muscle).…”

Section: Introductionmentioning

confidence: 99%

Intra- and intermuscular variations of postmortem protein degradation for PMI estimation

et al. 2020

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In recent years, protein decomposition has become of increasing interest for the use in forensic estimation of the postmortem interval (PMI). Especially skeletal muscle tissue has proven to be a prime target tissue, among other reasons, due to its large abundance in the human body. In this regard, it is important to know whether there are any intra-and intermuscular differences in the behavior of protein degradation. Thus, samples from different locations within several skeletal muscles as well as from cardiac and smooth muscle tissue samples were collected from three autopsy cases with varying degree of decomposition. Samples were analyzed by SDS-PAGE and Western blotting and compared for protein degradation patterns. Intramuscular variations turned out to be minimal and without major influence for the use of the method. Observed intermuscular differences provide possibilities for future improvement of the precision and temporal application range. The results of this study show the strengths and current limitations of protein degradation-based PMI estimation and provide a deeper understanding of intraindividual postmortem protein degradation processes.

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“…Saber and Ali (2016) showed a strong positive linear correlation between mRNA levels of Caspase-3 in the gastrocnemius muscle with time passed since death. They found a raise in mRNA levels of caspase-3 began 2 h postmortem until 6 h then a marked reduction occurred at 8 h. Lee et al (2016) has been reported that caspase-3 was completely degraded 84-96 h post-mortem (PMI 50 value = 26.3 h) in kidney by western blot and 76% of the stained area for caspase-3 was reduced gradually 48 h after death and totally absent at 96 h after death by immunohistochemistry. In another study, caspase levels started to decrease after 32 h post-mortem (Kemp et al, 2010).…”

Section: Discussionmentioning

confidence: 97%

Estimation of Postmortem Interval from mRNA Degradation and Autolytic Changes in the Brain and Adrenal Gland

Ali¹,

Hassan²,

Ramzy³

et al. 2018

Mansoura Journal of Forensic Medicine and Clinical Toxicology

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Estimation of the postmortem interval (PMI) is a very essential task for forensic experts especially in criminal cases. The PMI is the time passed since the death of an individual and refers to the stages of autolysis. This study aimed to estimate PMI from mRNA expression of Caspase-3 and IL-1β in the brain and adrenal gland of adult albino rats and the autolytic changes of these organs at time of death (0 hour), and then on 6, 12, 24, 48 hours postmortem (10 wistar male albino rats for each time). The results revealed a significant time-dependent elevation in the Caspase-3 mRNA expression levels of the brain and adrenal gland tissues, starting 6 h till 24 h postmortem then a significant reduction occurred at 48 h postmortem. On the other hand, the mRNA expression levels of IL-1β gene of the brain and adrenal gland tissues showed significant time-dependent reductions began at 6 h till 48 h postmortem. The results of the study concluded that the adrenal glands could be helpful for histopathological examination before 48 h at 20 °C and the brain tissue especially hippocampus could be helpful for histopathological examination before 12 h at 20 °C.

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Degradation of Kidney and Psoas Muscle Proteins as Indicators of Post-Mortem Interval in a Rat Model, with Use of Lateral Flow Technology

Cited by 28 publications

References 39 publications

Postmortem proteomics to discover biomarkers for forensic PMI estimation

Postmortem proteomics to discover biomarkers for forensic PMI estimation

Intra- and intermuscular variations of postmortem protein degradation for PMI estimation

Estimation of Postmortem Interval from mRNA Degradation and Autolytic Changes in the Brain and Adrenal Gland

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