Sachil Kumar1, Wahid Ali2, Uma Shankar Singh3, Ashutosh Kumar4, Sandeep Bhattacharya5, Anoop K Verma6. 1. Post Graduate Department of Pathology, King George's Medical University UP, Lucknow, India. Electronic address: sachilvohra@gmail.com. 2. Post Graduate Department of Pathology, King George's Medical University UP, Lucknow, India. Electronic address: aliwahid78@gmail.com. 3. Post Graduate Department of Pathology, King George's Medical University UP, Lucknow, India. Electronic address: ussinghjyotsna@yahoo.co.in. 4. Post Graduate Department of Pathology, King George's Medical University UP, Lucknow, India. Electronic address: ashutosh45kumar@gmail.com. 5. Department of Physiology, King George's Medical University UP, Lucknow, India. Electronic address: drsbhattacharya@gmail.com. 6. Department of Physiology, King George's Medical University UP, Lucknow, India. Electronic address: vermakgmc@gmail.com.
Abstract
BACKGROUND: After the death of an organism, intracellular enzymes cause protein to degrade into smaller fragments as the time passes, if these fragments can be isolated and visualized, and if the fragmentation is proved to be measurable and quantifiable, it can be a good sign of the post-mortem interval (PMI). The aim of this study is to evaluate the effect of PMI on Troponin-T protein degradation in cardiac tissues of cadavers through quantitative analysis of Troponin-T degradation by easily electrophoretic method and its association with PMI in case of deaths due to burn. METHODS: An efficient extraction protocol to analyze the banding pattern of cTnT in post-mortem tissue was developed. The analysis involves extraction of the protein (at different PMI (~7.30, 18.20, 30.30, 41.20, 41.40, 54.30..................88.40h), from the heart muscles of cadavers (burn cases), separation by denaturing gel electrophoresis (SDS-PAGE) and visualization by Western blot using cTnT specific monoclonal antibodies. RESULTS: The results specify a characteristic banding pattern amongst human cadavers (n=9), a pseudo-linear relationship between percent cTnT degraded and the time since death (r=0.87, p=0.0001). The area of the bands within a lane was quantified by scanning and digitizing the image using commonly available scanners. CONCLUSIONS: The present research used a simple, easy, applicable and highly informative electrophoresis method that make it an ideal for the busy forensic laboratory. The post-mortem Troponin-T fragmentation observed in this study reveals a sequential, time-dependent process with the potential for use as a predictor of PMI in cases of burning. This result shows a potential for use as a future applied method of evaluating time since death.
BACKGROUND: After the death of an organism, intracellular enzymes cause protein to degrade into smaller fragments as the time passes, if these fragments can be isolated and visualized, and if the fragmentation is proved to be measurable and quantifiable, it can be a good sign of the post-mortem interval (PMI). The aim of this study is to evaluate the effect of PMI on Troponin-T protein degradation in cardiac tissues of cadavers through quantitative analysis of Troponin-T degradation by easily electrophoretic method and its association with PMI in case of deaths due to burn. METHODS: An efficient extraction protocol to analyze the banding pattern of cTnT in post-mortem tissue was developed. The analysis involves extraction of the protein (at different PMI (~7.30, 18.20, 30.30, 41.20, 41.40, 54.30..................88.40h), from the heart muscles of cadavers (burn cases), separation by denaturing gel electrophoresis (SDS-PAGE) and visualization by Western blot using cTnT specific monoclonal antibodies. RESULTS: The results specify a characteristic banding pattern amongst human cadavers (n=9), a pseudo-linear relationship between percent cTnT degraded and the time since death (r=0.87, p=0.0001). The area of the bands within a lane was quantified by scanning and digitizing the image using commonly available scanners. CONCLUSIONS: The present research used a simple, easy, applicable and highly informative electrophoresis method that make it an ideal for the busy forensic laboratory. The post-mortem Troponin-T fragmentation observed in this study reveals a sequential, time-dependent process with the potential for use as a predictor of PMI in cases of burning. This result shows a potential for use as a future applied method of evaluating time since death.
Authors: Kyoung-Min Choi; Angela Zissler; Eunjung Kim; Bianca Ehrenfellner; Eunji Cho; Se-In Lee; Peter Steinbacher; Ki Na Yun; Jong Hwan Shin; Jin Young Kim; Walter Stoiber; Heesun Chung; Fabio Carlo Monticelli; Jae-Young Kim; Stefan Pittner Journal: Int J Legal Med Date: 2019-03-12 Impact factor: 2.686
Authors: Aniello Maiese; Andrea Scatena; Andrea Costantino; Marco Di Paolo; Raffaele La Russa; Emanuela Turillazzi; Paola Frati; Vittorio Fineschi Journal: Diagnostics (Basel) Date: 2021-01-03
Authors: Angela Zissler; Walter Stoiber; Peter Steinbacher; Janine Geissenberger; Fabio C Monticelli; Stefan Pittner Journal: Diagnostics (Basel) Date: 2020-11-26