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Post-mortem changes in metabolomic profiles of human serum, aqueous humor and vitreous humor

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Abstract

Introduction

Application of metabolomic methods to forensic studies may expand the limits of the post-mortem interval (PMI) estimation, and improve the accuracy of the estimation. To this end, it is important to determine which tissue is the most suitable for analysis, and which compounds are the most promising candidates for PMI estimation.

Objectives

This work is aimed at the comparison of human serum, aqueous humor (AH), and vitreous humor (VH) as perspective tissues for metabolomic-based PMI estimation, at the determination of most promising PMI biomarkers, and at the development of method of PMI estimation based on the measurement of concentrations of PMI biomarkers.

Methods

Quantitative metabolomic profiling of samples of the human serum, AH, and VH taken at different PMIs has been performed with the use of NMR spectroscopy.

Results

It is found that the metabolomic changes in anatomically isolated ocular fluids are slower and smoother than that in blood. A good positive time correlation (Pearson coefficient r > 0.5) was observed for several metabolites, including hypoxanthine, choline, creatine, betaine, glutamate, and glycine. A model for PMI estimation based on concentrations of several metabolites in AH and VH is proposed.

Conclusions

The obtained results demonstrate that the metabolomic analysis of AH and VH is more suitable for the PMI estimation than that of serum. The compounds with good positive time correlation can be considered as potential PMI biomarkers.

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References

  • Bartelink, E. J., & Chesson, L. A. (2019). Recent applications of isotope analysis to forensic anthropology. Forensic Sciences Research,4(1), 29–44.

    PubMed  PubMed Central  Google Scholar 

  • Donaldson, A. E., & Lamont, I. L. (2013). Biochemistry changes that occur after death: Potential markers for determining post-mortem interval. PLoS ONE,8(11), e82011.

    PubMed  PubMed Central  Google Scholar 

  • Donaldson, A. E., & Lamont, I. L. (2014). Estimation of post-mortem interval using biochemical markers. Australian Journal of Forensic Sciences,46(1), 8–26.

    Google Scholar 

  • Donaldson, A. E., & Lamont, I. L. (2015). Metabolomics of post-mortem blood: Identifying potential markers of post-mortem interval. Metabolomics,11(1), 237–245.

    CAS  Google Scholar 

  • Ferreira, P. G., Muñoz-Aguirre, M., Reverter, F., Sá Godinho, C. P., Sousa, A., Amadoz, A., et al. (2018). The effects of death and post-mortem cold ischemia on human tissue transcriptomes. Nature Communications,9(1), 490.

    PubMed  PubMed Central  Google Scholar 

  • Gonzalez-Riano, C., Tapia-González, S., García, A., Muñoz, A., DeFelipe, J., & Barbas, C. (2017). Metabolomics and neuroanatomical evaluation of post-mortem changes in the hippocampus. Brain Structure & Function,222(6), 2831–2853.

    CAS  Google Scholar 

  • Gowda, G. A., & Raftery, D. (2014). Quantitating metabolites in protein precipitated serum using NMR spectroscopy. Analytical Chemistry,86(11), 5433–5440.

    CAS  PubMed Central  Google Scholar 

  • Gowda, G. A., Gowda, Y. N., & Raftery, D. (2015). Expanding the limits of human blood metabolite quantitation using NMR spectroscopy. Analytical Chemistry,87(1), 706–715.

    PubMed  Google Scholar 

  • Gurler, M., Ozturk, G., Kir, M. Z., Ginis, Z., Erden, G., Akyol, S., et al. (2016). 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,48(2), 150–158.

    Google Scholar 

  • Henssge, C. (1988). Death time estimation in case work. I. The rectal temperature time of death nomogram. Forensic Science International,38(3–4), 209–236.

    CAS  PubMed  Google Scholar 

  • Henssge, C., Althaus, L., Bolt, J., Freislederer, A., Haffner, H. T., Henssge, C. A., et al. (2000). Experiences with a compound method for estimating the time since death. II. Integration of non-temperature-based methods. International Journal of Legal Medicine,113(6), 320–331.

    CAS  PubMed  Google Scholar 

  • Henssge, C., & Madea, B. (2004). Estimation of the time since death in the early post-mortem period. Forensic Science International,144(2–3), 167–175.

    CAS  PubMed  Google Scholar 

  • Henssge, C., Madea, B., & Gallenkemper, E. (1988). Death time estimation in case work. II. Integration of different methods. Forensic Science International,39(1), 77–87.

    CAS  PubMed  Google Scholar 

  • Henssge, C., & Madea, B. (2007). Estimation of the time since death. Forensic Science International,165(2–3), 182–184.

    PubMed  Google Scholar 

  • James, R. A., Hoadley, P. A., & Sampson, B. G. (1997). Determination of postmortem interval by sampling vitreous humour. The American Journal of Forensic Medicine and Pathology,18(2), 158–162.

    CAS  PubMed  Google Scholar 

  • Johnson, L. A., & Ferris, J. A. J. (2002). Analysis of postmortem DNA degradation by single-cell gel electrophoresis. Forensic Science International,126(1), 43–47.

    CAS  PubMed  Google Scholar 

  • Kryczka, T., Ehlers, N., Nielsen, K., Wylegala, E., Dobrowolski, D., & Midelfart, A. (2013). Metabolic profile of keratoconic cornea. Current Eye Research,38(2), 305–309.

    CAS  PubMed  Google Scholar 

  • Li, C., Ma, D., Deng, K., Chen, Y., Huang, P., & Wang, Z. (2017). Application of MALDI-TOF MS for estimating the postmortem interval in rat muscle samples. Journal of Forensic Sciences,62(5), 1345–1350.

    PubMed  Google Scholar 

  • Locci, E., Stocchero, M., Noto, A., Chighine, A., Natali, L., Napoli, P. E., et al. (2019). A 1H NMR metabolomic approach for the estimation of the time since death using aqueous humour: An animal model. Metabolomics: Official Journal of the Metabolomic Society,15(5), 76.

    Google Scholar 

  • Madea, B., Käferstein, H., Hermann, N., & Sticht, G. (1994). Hypoxanthine in vitreous humor and cerebrospinal fluid—a marker of postmortem interval and prolonged (vital) hypoxia? Remarks also on hypoxanthine in SIDS. Forensic Science International,65(1), 19–31.

    CAS  PubMed  Google Scholar 

  • Madea, B. (2016). Methods for determining time of death. Forensic Science, Medicine, and Pathology,12(4), 451–485.

    PubMed  Google Scholar 

  • Madea, B., & Rödig, A. (2006). Time of death dependent criteria in vitreous humor: Accuracy of estimating the time since death. Forensic Science International,164(2–3), 87–92.

    CAS  PubMed  Google Scholar 

  • Marshall, T. K., & Hoare, F. E. (1962). Estimating the time of death. The rectal cooling after death and its mathematical expression. Journal of Forensic Sciences,7, 56–81.

    Google Scholar 

  • Nazir, M. S., Smith, J. A., & Goodwin, W. (2011). DNA degradation in post-mortem soft muscle tissues in relation to accumulated degree-days (ADD). Forensic Science International: Genetics Supplement Series,3(1), e536–e537.

    Google Scholar 

  • Palmiere, C., & Mangin, P. (2015). Urea nitrogen, creatinine, and uric acid levels in postmortem serum, vitreous humor, and pericardial fluid. International Journal of Legal Medicine,129(2), 301–305.

    PubMed  Google Scholar 

  • Pittner, S., Ehrenfellner, B., Monticelli, F. C., Zissler, A., Sänger, A. M., Stoiber, W., et al. (2016). Postmortem muscle protein degradation in humans as a tool for PMI delimitation. International Journal of Legal Medicine,130(6), 1547–1555.

    PubMed  PubMed Central  Google Scholar 

  • Poulsen, J. P., Oyasaeter, S., Sanderud, J., Rognum, T. O., & Saugstad, O. D. (1990). Hypoxanthine, xanthine, and uric acid concentrations in the cerebrospinal fluid, plasma, and urine of hypoxemic pigs. Pediatric Research,28(5), 477–481.

    CAS  PubMed  Google Scholar 

  • Reibe, S., Doetinchem, P. V., & Madea, B. (2010). A new simulation-based model for calculating post-mortem intervals using developmental data for Lucilia sericata (Dipt.: Calliphoridae). Parasitology Research,107(1), 9–16.

    PubMed  Google Scholar 

  • Rognum, T. O., Hauge, S., Øyasaeter, S., & Saugstad, O. D. (1991). A new biochemical method for estimation of postmortem time. Forensic Science International,51(1), 139–146.

    CAS  PubMed  Google Scholar 

  • Rognum, T. O., Holmen, S., Musse, M. A., Dahlberg, P. S., Stray-Pedersen, A., Saugstad, O. D., et al. (2016). Estimation of time since death by vitreous humor hypoxanthine, potassium, and ambient temperature. Forensic Science International,262, 160–165.

    CAS  PubMed  Google Scholar 

  • Rosa, M. F., Scano, P., Noto, A., Nioi, M., Sanna, R., Paribello, F., et al. (2015). Monitoring the modifications of the vitreous humor metabolite profile after death: An animal model. BioMed Research International,2015, 627201.

    PubMed  PubMed Central  Google Scholar 

  • Sampaio-Silva, F., Magalhães, T., Carvalho, F., Dinis-Oliveira, R. J., & Silvestre, R. (2013). Profiling of RNA degradation for estimation of post mortem [corrected] interval. PLoS ONE,8(2), e56507.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sato, T., Zaitsu, K., Tsuboi, K., Nomura, M., Kusano, M., Shima, N., et al. (2015). A preliminary study on postmortem interval estimation of suffocated rats by GC-MS/MS-based plasma metabolic profiling. Analytical and Bioanalytical Chemistry,407(13), 3659–3665.

    CAS  PubMed  Google Scholar 

  • Sharma, R., Kumar Garg, R., & Gaur, J. R. (2015). Various methods for the estimation of the post mortem interval from Calliphoridae: A review. Egyptian Journal of Forensic Sciences,5(1), 1–12.

    Google Scholar 

  • Snytnikova, O. A., Khlichkina, A. A., Yanshole, L. V., Yanshole, V. V., Iskakov, I. A., Egorova, E. V., et al. (2016). Metabolomics of the human aqueous humor. Metabolomics,13(1), 5.

    Google Scholar 

  • Snytnikova, O. A., Yanshole, L. V., Iskakov, I. A., Yanshole, V. V., Chernykh, V. V., Stepakov, D. A., et al. (2017). Quantitative metabolomic analysis of the human cornea and aqueous humor. Metabolomics,13(12), 152.

    Google Scholar 

  • Streete, I. M., Jamie, J. F., & Truscott, R. J. W. (2004). Lenticular levels of amino acids and free UV filters differ significantly between normals and cataract patients. Investigative Ophthalmology & Visual Science,45(11), 4091–4098.

    Google Scholar 

  • Swain, R., Kumar, A., Sahoo, J., Lakshmy, R., Gupta, S. K., Bhardwaj, D. N., et al. (2015). Estimation of post-mortem interval: A comparison between cerebrospinal fluid and vitreous humour chemistry. Journal of Forensic and Legal Medicine,36, 144–148.

    PubMed  Google Scholar 

  • Takata, T., Kitao, T., & Miyaishi, S. (2014). Relationship between post-mortem interval and creatine concentration in vitreous humour and cerebrospinal fluid. Australian Journal of Forensic Sciences,46(2), 160–165.

    Google Scholar 

  • Tsentalovich, Y. P., Verkhovod, T. D., Yanshole, V. V., Kiryutin, A. S., Yanshole, L. V., Fursova, A. Z., et al. (2015). Metabolomic composition of normal aged and cataractous human lenses. Experimental Eye Research,134, 15–23.

    CAS  PubMed  Google Scholar 

  • Tumram, N. K., Bardale, R. V., & Dongre, A. P. (2011). Postmortem analysis of synovial fluid and vitreous humour for determination of death interval: A comparative study. Forensic Science International,204(1–3), 186–190.

    CAS  PubMed  Google Scholar 

  • Verica, P., Janeska, B., Gutevska, A., & Duma, A. (2007). Post mortem cooling of the body and estimation of time since death. Soudni Lekarstvi,52(4), 50–56.

    PubMed  Google Scholar 

  • Yanshole, V. V., Yanshole, L. V., Snytnikova, O. A., & Tsentalovich, Y. P. (2019). Quantitative metabolomic analysis of changes in the lens and aqueous humor under development of age-related nuclear cataract. Metabolomics,15(3), 29.

    PubMed  Google Scholar 

  • Zelentsova, E. A., Yanshole, L. V., Snytnikova, O. A., Yanshole, V. V., Tsentalovich, Y. P., & Sagdeev, R. Z. (2016). Post-mortem changes in the metabolomic compositions of rabbit blood, aqueous and vitreous humors. Metabolomics,12(11), 172.

    Google Scholar 

  • Zelentsova, E. A., Yanshole, V. V., & Tsentalovich, Y. P. (2019). A novel method of sample homogenization with the use of a microtome-cryostat apparatus. RSC Advances,9(65), 37809–37817.

    CAS  Google Scholar 

  • Zhu, Y., Wang, L., Yin, Y., & Yang, E. (2017). Systematic analysis of gene expression patterns associated with postmortem interval in human tissues. Scientific Reports,7(1), 1–12.

    Google Scholar 

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Funding

This research was funded by Russian Foundation for Basic Research (Project 18-33-20097 in NMR measurements, and Project 18-29-13023 in sample preparation). We thank Ministry of Science and Higher Education of the RF for the access to NMR equipment.

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Contributions

IK and VN collected samples. EZ and YT conceived and designed research. EZ and LY conducted experiments. AM analyzed data. YP wrote the manuscript and supervised the research. All authors read and approved the manuscript.

Corresponding author

Correspondence to Yuri P. Tsentalovich.

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All authors declare that they do not have conflict of interest.

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All procedures of this study were subjected to the Declaration of Helsinki—ethical principles for medical research involving human subjects and with the ethical approval from International Tomography Center SB RAS (ECITC-2017-03).

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Zelentsova, E.A., Yanshole, L.V., Melnikov, A.D. et al. Post-mortem changes in metabolomic profiles of human serum, aqueous humor and vitreous humor. Metabolomics 16, 80 (2020). https://doi.org/10.1007/s11306-020-01700-3

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