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The Use of Molecular Markers in Forensic Field as an Element of Scientific Evidence

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Biology".

Deadline for manuscript submissions: 20 August 2025 | Viewed by 7606

Special Issue Editors

Dr. Angelo Montana

E-Mail Website
Guest Editor
Department of Excellence Biomedical Sciences and Public Health, Marche Polytechnic University, 60121 Ancona, Italy
Interests: forensic toxicology; forensic pathology; organ damage, drug toxicity forensic biomarkers
Special Issues, Collections and Topics in MDPI journals
Dr. Margherita Neri

E-Mail Website
Guest Editor
Department of Medical Sciences, University of Ferrara, 44121 Ferrara, Italy
Interests: forensic toxicology; forensic pathology; organ damage; drug toxicity; forensic biomarkers

Special Issue Information

Dear Colleagues,

In recent years, research on the role of molecular markers has had a significant increase in the forensic field. Biochemical markers are analyzed in the blood, urine, cerebrospinal fluid, or other biological samples. They may provide information about the cause of death and the post-mortem interval (PMI). The forensic molecular pathology procedure can be integrated into routine work to improve and to reinforce morphological evidence. It could be applied in medical sciences to investigate the pathophysiology of the diseases and trauma that led to death. For example, the immunohistochemical characteristics of vitality in hanging and the identification of the most significant vitality markers on ligature marks could be useful to determine whether the hanging was committed as suicide or as a simulated hanging. For tissue identification in a forensic context, it is possible to employ many molecular markers, such as mRNA, miRNA, DNA methylation, and microbial markers. Biochemical markers, using a femoral venous blood sample, can also detect cerebral damage and acute phase response in early post-mortem through measurements of GFAP, NSE, and BDNF. In the context of molecular pathology, the subjects include: postmortem interval determination via miRNAs, mRNAs, or microbial markers and cause of death by cardiac disease determined via mRNA markers or by drowning through microbial markers. Microbial markers could also help to mark a fatal hospital infection or to compare ante-mortem burns and post-mortem burns using an mRNA marker. The objective of this Special Issue is to collect original and review articles to provide several markers for forensic pathologists to be used as scientific evidence for forensic research and justice.

Dr. Angelo Montana
Dr. Margherita Neri
Guest Editors

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Keywords

  • biomarkers
  • immunohistochemistry
  • histopathology
  • forensic pathology
  • forensic toxicology
  • forensic biomarkers
  • toxicological biomarkers
  • neuropathology
  • mRNA
  • post-mortem interval
  • cerebral damage
  • in vitro, in vivo, and in silico models
  • hypoxic–ischemic brain injury
  • pregnancy biomarkers
  • vitality

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Published Papers (3 papers)

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Research

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25 pages, 4309 KiB  
Article
Development of Mathematical Models Using circRNA Combinations (circTulp4, circSlc8a1, and circStrn3) in Mouse Brain Tissue for Postmortem Interval Estimation
by Binghui Song, Jiewen Fu, Jie Qian, Ting He, Jingliang Cheng, Sawitree Chiampanichayakul, Songyot Anuchapreeda and Junjiang Fu
Int. J. Mol. Sci. 2025, 26(10), 4495; https://doi.org/10.3390/ijms26104495 - 8 May 2025
Viewed by 423
Abstract
The postmortem interval (PMI) is defined as the time interval between physiological death and the examination of the corpse, playing a critical role in forensic investigations. Traditional PMI estimation methods are often influenced by subjective and environmental factors. Circular RNAs (circRNAs), known for [...] Read more.
The postmortem interval (PMI) is defined as the time interval between physiological death and the examination of the corpse, playing a critical role in forensic investigations. Traditional PMI estimation methods are often influenced by subjective and environmental factors. Circular RNAs (circRNAs), known for their stability, abundance, and conservation in brain tissue, show promise as biomarkers for PMI estimation. However, research on circRNAs in this context remains limited. This study aimed to develop PMI estimation models using circRNAs across multiple temperatures. By employing semi-quantitative reverse transcription-PCR, circTulp4, circSlc8a1, and circStrn3 were identified as reliable biomarkers for mouse brain tissue. Mathematical models were constructed using the reference genes 28S rRNA, mt-co1, and circCDR1as. At 4 °C, most equations had p-values below 0.05, with the equation using circSlc8a1 as a marker exhibiting the highest goodness of fit. Validation results indicated that the equation using circTulp4 as the reference gene had the highest accuracy. When applying the combined aforementioned three circRNAs, the equation using circCDR1as as the reference gene showed better accuracy. At 25 °C, all equations had R2 values greater than 0.86, but most cubic equations had p-values above 0.05. Validation results demonstrated that the circTulp4/mt-co1 equation had the highest accuracy. When applying combined circRNAs, the R2 values improved, and long-term PMI estimation was more accurate than short-term PMI estimation. At 35 °C, the linear equations had significantly poorer goodness of fit compared to nonlinear equations, and nonlinear equations exhibited better accuracy than linear equations. When applying the combined aforementioned three circRNAs, the accuracy of the three reference genes was similar, and the accuracy of long-term PMI estimation was consistently higher than that of short-term estimation. For the three-dimensional models, all R2 values exceeded 0.75 with p-values significantly below 0.0001. Validation results demonstrated higher accuracy at 25 °C and 35 °C, with superior performance for long-term PMI estimation. In summary, this study constructed PMI estimation models under multiple temperature conditions based on highly expressed circRNAs in mouse brain tissue, highlighting circTulp4, circSlc8a1, and circStrn3 as novel biomarkers. These findings offer a complementary tool for PMI estimation, particularly for long-term PMI estimation. Full article
(This article belongs to the Special Issue The Use of Molecular Markers in Forensic Field as an Element of Scientific Evidence)
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13 pages, 1124 KiB  
Article
In Vitro and In Vivo Human Metabolism of Ostarine, a Selective Androgen Receptor Modulator and Doping Agent
by Omayema Taoussi, Giulia Bambagiotti, Prince Sellase Gameli, Gloria Daziani, Francesco Tavoletta, Anastasio Tini, Giuseppe Basile, Alfredo Fabrizio Lo Faro and Jeremy Carlier
Int. J. Mol. Sci. 2024, 25(14), 7807; https://doi.org/10.3390/ijms25147807 - 17 Jul 2024
Cited by 4 | Viewed by 4615
Abstract
Ostarine (enobasarm) is a selective androgen receptor modulator with great therapeutic potential. However, it is also used by athletes to promote muscle growth and enhance performances without the typical adverse effects of anabolic steroids. Ostarine popularity increased in recent years, and it is [...] Read more.
Ostarine (enobasarm) is a selective androgen receptor modulator with great therapeutic potential. However, it is also used by athletes to promote muscle growth and enhance performances without the typical adverse effects of anabolic steroids. Ostarine popularity increased in recent years, and it is currently the most abused “other anabolic agent” (subclass S1.2. of the “anabolic agents” class S1) from the World Anti-Doping Agency’s (WADA) prohibited list. Several cases of liver toxicity were recently reported in regular users. Detecting ostarine or markers of intake in biological matrices is essential to document ostarine use in doping. Therefore, we sought to investigate ostarine metabolism to identify optimal markers of consumption. The substance was incubated with human hepatocytes, and urine samples from six ostarine-positive cases were screened. Analyses were performed via liquid chromatography–high-resolution tandem mass spectrometry (LC-HRMS/MS) and software-assisted data mining, with in silico metabolite predictions. Ten metabolites were identified with hydroxylation, ether cleavage, dealkylation, O-glucuronidation, and/or sulfation. The production of cyanophenol-sulfate might participate in the mechanism of ostarine liver toxicity. We suggest ostarine-glucuronide (C25H22O9N3F3, diagnostic fragments at m/z 118, 185, and 269) and hydroxybenzonitrile-ostarine-glucuronide (C25H22O10N3F3, diagnostic fragments at m/z 134, 185, and 269) in non-hydrolyzed urine and ostarine and hydroxybenzonitrile-ostarine (C19H14O4N3F3, diagnostic fragments at m/z 134, 185, and 269) in hydrolyzed urine as markers to document ostarine intake in doping. Full article
(This article belongs to the Special Issue The Use of Molecular Markers in Forensic Field as an Element of Scientific Evidence)
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Review

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22 pages, 597 KiB  
Review
Potential Role of mRNA in Estimating Postmortem Interval: A Systematic Review
by Vincenzo Cianci, Cristina Mondello, Daniela Sapienza, Maria Cristina Guerrera, Alessio Cianci, Annalisa Cracò, Fausto Omero, Vittorio Gioffrè, Patrizia Gualniera, Alessio Asmundo and Antonino Germanà
Int. J. Mol. Sci. 2024, 25(15), 8185; https://doi.org/10.3390/ijms25158185 - 26 Jul 2024
Cited by 3 | Viewed by 1554
Abstract
Although the postmortem interval estimation still represents one of the main goals of forensic medicine, there are still several limitations that weigh on the methods most used for its determination: for this reason, even today, precisely estimating the postmortem interval remains one of [...] Read more.
Although the postmortem interval estimation still represents one of the main goals of forensic medicine, there are still several limitations that weigh on the methods most used for its determination: for this reason, even today, precisely estimating the postmortem interval remains one of the most important challenges in the forensic pathology field. To try to overcome these limitations, in recent years, numerous studies have been conducted on the potential use of the mRNA degradation time for reaching a more precise post mortem interval (PMI) estimation. An evidence-based systematic review of the literature has been conducted to evaluate the state of the art of the knowledge focusing on the potential correlation between mRNA degradation and PMI estimation. The research has been performed using the electronic databases PubMed and Scopus. The analysis conducted made it possible to confirm the potential applicability of mRNA for reaching a more precise PMI estimation. The analysis of the results highlighted the usefulness of some mRNAs, such as β-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA, especially in short time frames, within a few hours or days of death. The matrices on which these analyses were conducted were also analyzed, resulting in less exposure to the external environment, including the heart, brain, and dental pulp. The major limitations were also reported, including the short time intervals analyzed in most of the articles, the lack of mathematical models, and the failure to report the error rate between the mRNA degradation time and PMI. Given the still small number of published articles, the lack of globally recognized standardized methods, and the numerous techniques used to evaluate the mRNA degradation times, numerous and larger studies are still necessary to reach more solid and shared evidence. Full article
(This article belongs to the Special Issue The Use of Molecular Markers in Forensic Field as an Element of Scientific Evidence)
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