POST-MORTEM CHANGES IN METABOLOMIC PROFILES OF HUMAN SERUM, AQUEOUS HUMOR, AND VITREOUS HUMOR

L RIAZ; AM SIDDIQUI; MN RASHID; RA SHAHID; . NASRULHUDA; F BUTT

doi:10.54112/bcsrj.v2024i1.966

Authors

L RIAZ Department of Forensic Medicine, Dow Medical College, Dow University of Health Sciences, Karachi, Pakistan
AM SIDDIQUI Department of Anatomy, Dow University of Health Sciences Karachi, Pakistan
MN RASHID Department of Physiology, Director Postgraduate Medical Education, Research and Ethics Department, Shaheed Mohtarma Benazir Bhutto Medical College, Dow University of Health Sciences, Karachi, Pakistan
RA SHAHID Department of Physiology, Dow University of Health Sciences Karachi, Pakistan
. NASRULHUDA Department of Anatomy, Dow University of Health Sciences Karachi, Pakistan
F BUTT Department of Physiology, Dow University of Health Sciences Karachi, Pakistan

DOI:

https://doi.org/10.54112/bcsrj.v2024i1.966

Keywords:

Metabolomics, Post-Mortem, Aqueous Humor, Vitreous Humor.

Abstract

Accurate post-mortem interval (PMI) estimation is crucial in forensic investigations, and metabolomic analysis of bodily fluids offers a promising approach. However, the suitability of ocular fluids for PMI estimation remains underexplored. Objective: The objective of this study was to evaluate the suitability of metabolomic analysis of ocular fluids, specifically vitreous humor (VH) and aqueous humor (AH), compared to serum for post-mortem interval (PMI) estimation and to identify the most promising metabolites for this purpose. Methods: A cross-sectional study was conducted at a tertiary care hospital in Karachi. Blood, AH, and VH samples were collected from human cadavers with PMIs ranging from 5 to 60 hours. Samples were prepared by removing proteins and lipids, followed by lyophilization. Metabolomic profiling was performed using an NMR Spectrometer. Pearson correlation coefficients were calculated to assess the relationship between metabolite concentrations and PMI. Results: The study identified 24 metabolites in blood, AH, and VH samples. Several metabolites, including glutamate, choline, glycine, and formate, exhibited significant positive correlations with PMI in all three fluids, with Pearson coefficients greater than 0.5. Notably, glutamate (blood: r = 0.585**, AH: r = 0.488*, VH: r = 0.683**), choline (blood: r = 0.435*, AH: r = 0.691**, VH: r = 0.713**), and glycine (blood: r = 0.553*, AH: r = 0.598**, VH: r = 0.690**) showed strong correlations. Conversely, glucose and pyruvate significantly negatively correlated with PMI in AH and VH. Conclusion: The findings indicate that VH and AH are more reliable than serum for PMI estimation due to their more stable metabolite profiles. Glutamate, choline, and glycine emerged as particularly promising biomarkers. The study supports the potential of metabolomic analysis of ocular fluids in forensic investigations to provide accurate PMI estimations.

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References

Wolstenholme R, Jickells S, Forbes S. Analytical techniques in forensic science: John Wiley & Sons; 2021.

Żarczyńska M, Żarczyński P, Tomsia M. Nucleic acids persistence—benefits and limitations in forensic genetics. Genes. 2023;14(8):1643.

Cláudia-Ferreira A, Barbosa DJ, Saegeman V, Fernández-Rodríguez A, Dinis-Oliveira RJ, Freitas AR, et al. The future is now: unraveling the expanding potential of human (Necro) microbiome in forensic investigations. Microorganisms. 2023;11(10):2509.

Locci E, Stocchero M, Gottardo R, De-Giorgio F, Demontis R, Nioi M, et al. Comparative use of aqueous humour 1H NMR metabolomics and potassium concentration for PMI estimation in an animal model. International Journal of Legal Medicine. 2021;135(3):845-52.

Pigaiani N, Bertaso A, De Palo EF, Bortolotti F, Tagliaro F. Vitreous humor endogenous compounds analysis for post-mortem forensic investigation. Forensic science international. 2020;310:110235.

Locci E, Stocchero M, Gottardo R, Chighine A, De-Giorgio F, Ferino G, et al. PMI estimation through metabolomics and potassium analysis on animal vitreous humour. International Journal of Legal Medicine. 2023;137(3):887-95.

Zelentsova EA, Yanshole LV, Melnikov AD, Kudryavtsev IS, Novoselov VP, Tsentalovich YP. Post-mortem changes in metabolomic profiles of human serum, aqueous humor and vitreous humor. Metabolomics. 2020;16:1-10.

Gurler M, Ozturk G, Kir MZ, Ginis Z, Erden G, Akyol S, et al. Simultaneous analysis of biochemical markers in vitreous humour and serum: a preliminary study on the effect of storage time at–20° C. Australian Journal of Forensic Sciences. 2016;48(2):150-8.

Zelentsova EA, Yanshole LV, Snytnikova OA, Yanshole VV, Tsentalovich YP, Sagdeev RZ. Post-mortem changes in the metabolomic compositions of rabbit blood, aqueous and vitreous humors. Metabolomics. 2016;12:1-11.

Steuer AE, Wartmann Y, Schellenberg R, Mantinieks D, Glowacki LL, Gerostamoulos D, et al. Postmortem metabolomics: influence of time since death on the level of endogenous compounds in human femoral blood. Necessary to be considered in metabolome study planning? Metabolomics. 2024;20(3):51.

Snytnikova OA, Yanshole LV, Iskakov IA, Yanshole VV, Chernykh VV, Stepakov DA, et al. Quantitative metabolomic analysis of the human cornea and aqueous humor. Metabolomics. 2017;13:1-9.

Fayyaz A. Experimental and computational ocular pharmacokinetics with cocktail administration for ocular drug development: Itä-Suomen yliopisto; 2022.

Yanshole VV, Yanshole LV, Snytnikova OA, Tsentalovich YP. Quantitative metabolomic analysis of changes in the lens and aqueous humor under development of age-related nuclear cataract. Metabolomics. 2019;15:1-11.

Rognum T, Holmen S, Musse M, Dahlberg P, Stray-Pedersen A, Saugstad O, et al. Estimation of time since death by vitreous humor hypoxanthine, potassium, and ambient temperature. Forensic science international. 2016;262:160-5.

Locci E, Stocchero M, Noto A, Chighine A, Natali L, Napoli PE, et al. A 1 H NMR metabolomic approach for the estimation of the time since death using aqueous humour: an animal model. Metabolomics. 2019;15:1-13.