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Analysis of immunohistochemical expression of inducible nitric oxide synthase for the evaluation of agonal time in forensic medicine

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Abstract

Although establishing agony is crucial in forensic practice, the identification of specific signs indicative of a detailed duration of agony is however not of immediate execution. Nitric oxide (NO) is the most important messenger molecule in the modulation of vascular tone and it is produced during stress conditions by inducible nitric oxide synthase (iNOS), as occurs during agony. The aim of this study was to investigate the relationship between immunohistochemical expression of iNOS, and agonal time (T), defined as the interval between the onset of a hypoxic-ischemic injury and the death. INOS expression was evaluated by measuring the average of signal intensity (SI) from cytoplasm of 300 smooth muscle cells of sample of renal artery, performed by ImageJ software: high values of SI correspond to a low enzyme expression and vice versa. We aimed also to check if gender, age, type of death (violent or natural death), post mortem interval, and storage in cold chamber influenced SI. We assessed 50 autopsied cases, of which 28 violent and 22 natural deaths, with a well-known T in a range between 1 and 631 min. Statistical analysis was performed to estimate the relationship between SI and the other variables. Results pointed out that only SI is related to T, and since data showed a bi-phase relationship between T and SI, we used a piecewise regression method for estimation of T as function of SI. The transition from the first to the second phase takes place at SI = 117.5 which corresponds to a T of 29.5 min. In conclusion, the study demonstrates that iNOS is a good marker for estimating T and the final regression model can be used in many forensic activities.

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Acknowledgments

This work was supported by grants from Polytechnic University of Marches. We will like to thank all members of Section of Legal Medicine of Polytechnic University of Marches for providing the samples of tissues.

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Author notes

  1. R. Scendoni

    Present address: Institute of Legal Medicine, Department of Law, University of Macerata, Macerata, 62100, Italy

Authors and Affiliations

  1. Department of Biomedical Sciences and Public Health, Section of Legal Medicine, Faculty of Medicine, Polytechnic University of Marche, Torrette di Ancona, 60020, Italy

    R. Scendoni & A. Tagliabracci

  2. Center of Epidemiology, Biostatistics and Medical Information Technology, Department of Biomedical Sciences and Public Health, Polytechnic University of Marche, Torrette di Ancona, 60020, Italy

    L. Ferrante

  3. Department of Biomedical Sciences and Public Health, Section of Pathological Anatomy, Polytechnic University of Marche, Torrette di Ancona, 60020, Italy

    D. Stramazzotti

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  1. R. Scendoni
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  2. L. Ferrante
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  3. D. Stramazzotti
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  4. A. Tagliabracci
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Corresponding author

Correspondence to A. Tagliabracci.

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Our investigations were conducted in accordance with the human and ethical principles of our university.

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The authors declare that they have no conflicts of interest.

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Scendoni, R., Ferrante, L., Stramazzotti, D. et al. Analysis of immunohistochemical expression of inducible nitric oxide synthase for the evaluation of agonal time in forensic medicine. Int J Legal Med 130, 1639–1646 (2016). https://doi.org/10.1007/s00414-016-1402-8

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  • DOI: https://doi.org/10.1007/s00414-016-1402-8

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