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Molecular Biology in Forensic Science: Past, Present and Future

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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: closed (31 December 2023) | Viewed by 21888

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Dr. Francesco Sessa

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Department of Medical, Surgical and Advanced Technologies “G.F. Ingrassia”, University of Catania, 95121 Catania, Italy
Interests: genetics and molecular biology; forensic and biological sciences; forensic genetics; genomic physiology; aging and genetics; pharmacology; toxicology; health professions; translational pharmacology; biochemistry
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Dr. Monica Salerno

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Guest Editor

Department of Medical, Surgical and Advanced Technologies “G.F. Ingrassia”, University of Catania, 95121 Catania, Italy
Interests: forensic sciences; clinical and forensic autopsy; crime scene investigation; post-mortem investigation; forensic pathology; histopathology; immunohistochemistry; medical risk management; ethics
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Molecular biology represents one of the most valuable tools currently used in bioscience, diagnostics and forensic sciences. It is important to remark that when considering the current and future trends in forensic molecular biology it is important to do so not only from the perspective of their effect within the forensic field itself, but also from the perspective of their interaction with neighboring areas of the system, such as the diagnostic field. Forensic molecular biology has developed rapidly into a comprehensive discipline in its own right and, perhaps more so than any scientific advance before it, has had a profound impact across the forensic sciences. For example, forensic DNA analysis has vastly evolved since the first forensic samples were evaluated by restriction fragment length polymorphism (RFLP). Methodologies advanced from gel electrophoresis techniques to capillary electrophoresis, and now to next-generation sequencing (NGS). NGS is the newest and most revolutionary technology, and has the potential to be the next standard for genetic analysis. In the same way, applications related to the use of microRNAs (miRNAs) represent a fascinating research field for both forensic and healthcare purposes.

In this regard, this Special Issue aims to collect accurate and up-to-date scientific information on all aspects of forensic molecular biology techniques and applications that are currently used for the analysis of human and nonhuman samples, in the form of original research articles, case series and case reports, and reviews.

Dr. Francesco Sessa
Dr. Monica Salerno
Guest Editors

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Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

genetics and molecular biology
forensic molecular biology
forensic and biological sciences
forensic genetics
aging and genetics
molecular biology and pharmacology
molecular biology and toxicology
translational pharmacology
biochemistry

Related Special Issue

Molecular Biology in Forensic Science: Past, Present and Future 2.0 in International Journal of Molecular Sciences (5 articles)

Published Papers (11 papers)

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Editorial

Jump to: Research, Review

6 pages, 185 KiB

Open AccessEditorial

Special Issue “Molecular Biology in Forensic Science: Past, Present and Future”

by Francesco Sessa and Monica Salerno

Int. J. Mol. Sci. 2024, 25(5), 2883; https://doi.org/10.3390/ijms25052883 - 01 Mar 2024

Viewed by 764

Abstract

Molecular biology has always represented an enviable tool in the fields of biosciences, diagnostics, and forensic sciences [...] Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

Research

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21 pages, 932 KiB

Open AccessArticle

Trace DNA Transfer in Co-Working Spaces: The Importance of Background DNA Analysis

by Martina Onofri, Federica Tommolini, Simona Severini, Cristiana Gambelunghe, Massimo Lancia, Luigi Carlini and Eugenia Carnevali

Int. J. Mol. Sci. 2024, 25(4), 2207; https://doi.org/10.3390/ijms25042207 - 12 Feb 2024

Viewed by 664

Abstract

The presence of background DNA (bgDNA) can hinder the evaluation of DNA evidence at the activity level, especially when the suspect is expected to be retrieved due to their habitual occupation of the investigated environment. Based on real-life casework circumstances, this study investigates the prevalence, composition, origin, and probable transfer routes of bgDNA found on personal items in situations where their owner and person of interest (POI) share the same workspace. Baseline values of bgDNA were evaluated on the participants’ personal items. Secondary and higher degree transfer scenarios of non-self DNA deposition were also investigated. The DNA from co-workers and co-inhabiting partners can be recovered from an individual’s personal belongings. Non-self DNA present on the hands and deposited on a sterile surface can generate uninformative profiles. The accumulation of foreign DNA on surfaces over time appears to be crucial for the recovery of comparable profiles, resulting in detectable further transfer onto other surfaces. For a thorough evaluation of touch DNA traces at the activity level, it is necessary to collect information not only about DNA transfer probabilities but also about the presence of the POI as part of the ‘baseline’ bgDNA of the substrates involved. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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10 pages, 941 KiB

Open AccessCommunication

Forensic Proteomics for the Discovery of New post mortem Interval Biomarkers: A Preliminary Study

by Alessandro Marrone, Daniele La Russa, Laura Barberio, Maria Stella Murfuni, Marco Gaspari and Daniela Pellegrino

Int. J. Mol. Sci. 2023, 24(19), 14627; https://doi.org/10.3390/ijms241914627 - 27 Sep 2023

Cited by 1 | Viewed by 1138

Abstract

Estimating the time since death (post mortem interval, PMI) represents one of the most important tasks in daily forensic casework. For decades, forensic scientists have investigated changes in post mortem body composition, focusing on different physical, chemical, or biological aspects, to discover a reliable method for estimating PMI; nevertheless, all of these attempts remain unsuccessful considering the currently available methodical spectrum characterized by great inaccuracies and limitations. However, recent promising approaches focus on the post mortem decomposition of biomolecules. In particular, significant advances have been made in research on the post mortem degradation of proteins. In the present study, we investigated early post mortem changes (during the first 24 h) in the proteome profile of the pig skeletal muscle looking for new PMI specific biomarkers. By mass spectrometry (MS)-based proteomics, we were able to identify a total of nine potential PMI biomarkers, whose quantity changed constantly and progressively over time, directly or inversely proportional to the advancement of post mortem hours. Our preliminary study underlines the importance of the proteomic approach in the search for a reliable method for PMI determination and highlights the need to characterize a large number of reliable marker proteins useful in forensic practice for PMI estimation. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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13 pages, 3666 KiB

Open AccessArticle

Partners in Postmortem Interval Estimation: X-ray Diffraction and Fourier Transform Spectroscopy

by Leticia Rubio, Juan Suárez, Stella Martin-de-las-Heras and Sara C. Zapico

Int. J. Mol. Sci. 2023, 24(7), 6793; https://doi.org/10.3390/ijms24076793 - 05 Apr 2023

Cited by 3 | Viewed by 1883

Abstract

The postmortem interval (PMI) is difficult to estimate in later stages of decomposition. There is therefore a need to develop reliable methodologies to estimate late PMI. This study aims to assess whether there is a correlation between changes in the mineral composition of human teeth and the estimation of PMI. X-ray diffraction (XRD) and attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy techniques were performed to address this challenge. Forty healthy human teeth obtained from odontological clinics were stored at different times (0, 10, 25, 50 years; N = 10/group). XRD and ATR-FTIR parameters related to the structure and composition of teeth were studied. Our results showed that the crystallinity index, crystal size index, mineral-to-organic matrix ratio (M/M) and carbonate/phosphate ratio (C/P) had the strongest association with PMI. For larger PMIs, there was a significant increase in crystallinity, crystal size and M/M ratio, while the C/P ratio showed a specific decrease with increasing PMI. According to our results, the parameters of crystallinity, crystal size, M/M ratio and C/P ratio can be considered highly accurate in determining a PMI of 10 years of data; crystallinity and mineral maturity can be considered useful in determining a PMI of 25 years; and crystallinity and mineral maturity can be considered highly accurate in determining a PMI of 50 years. A particular XRD index was identified as the most suitable parameter to estimate PMI: crystallinity. The joint use of XRD and ATR-FTIR analyses could be a promising alternative for dating human teeth. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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21 pages, 9921 KiB

Open AccessArticle

New Insight into Mechanisms of Cardiovascular Diseases: An Integrative Analysis Approach to Identify TheranoMiRNAs

by Francesco Sessa, Monica Salerno, Massimiliano Esposito, Giuseppe Cocimano, Daniela Pisanelli, Abdul Malik, Azmat Ali Khan and Cristoforo Pomara

Int. J. Mol. Sci. 2023, 24(7), 6781; https://doi.org/10.3390/ijms24076781 - 05 Apr 2023

Cited by 2 | Viewed by 1936

Abstract

MiRNAs regulate both physiological and pathological heart functions. Altered expression of miRNAs is associated with cardiovascular diseases (CVDs), making miRNAs attractive therapeutic strategies for the diagnosis and treatment of heart diseases. A recent publication defined, for the first time, the term theranoMiRNA, meaning the miRNAs that may be used both for diagnosis and treatment. The use of in silico tools may be considered fundamental for these purposes, clarifying several molecular aspects, suggesting future directions for in vivo studies. This study aims to explore different bioinformatic tools in order to clarify miRNA interactions with candidate genes, demonstrating the need to use a computational approach when establishing the most probable associations between miRNAs and target genes. This study focused on the functions of miR-133a-3p, miR-21-5p, miR-499a-5p, miR-1-3p, and miR-126-3p, providing an up-to-date overview, and suggests future lines of research in the identification of theranoMiRNAs related to CVDs. Based on the results of the present study, we elucidated the molecular mechanisms that could be linked between miRNAs and CVDs, confirming that these miRNAs play an active role in the genesis and development of heart damage. Given that CVDs are the leading cause of death in the world, the identification of theranoMiRNAs is crucial, hence the need for a definition of in vivo studies in order to obtain further evidence in this challenging field of research. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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13 pages, 1265 KiB

Open AccessArticle

Forensic Age Estimation through a DNA Methylation-Based Age Prediction Model in the Italian Population: A Pilot Study

by Martina Onofri, Arianna Delicati, Beatrice Marcante, Luigi Carlini, Federica Alessandrini, Pamela Tozzo and Eugenia Carnevali

Int. J. Mol. Sci. 2023, 24(6), 5381; https://doi.org/10.3390/ijms24065381 - 11 Mar 2023

Cited by 4 | Viewed by 1815

Abstract

DNA methylation is one of the epigenetic marks which has been studied intensively in recent years for age predicting purposes in the forensic area. In order to integrate age prediction into routine forensic workflow, the purpose of this study was to standardize and optimize a DNA methylation-based protocol tailored to the Italian context. A previously published protocol and age-predictive method was implemented for the analysis of 84 blood samples originating from Central Italy. The study here presented is based on the Single Base Extension method, considering five genes: ELOVL2, FHL2, KLF14, C1orf132, now identified as MIR29B2C, and TRIM59. The precise and specific steps consist of DNA extraction and quantification, bisulfite conversion, amplification of converted DNA, first purification, single base extension, second purification, capillary electrophoresis, and analysis of the results to train and test the tool. The prediction error obtained, expressed as mean absolute deviation, showed a value of 3.12 years in the training set and 3.01 years in the test set. Given that population-based differences in DNA methylation patterns have been previously reported in the literature, it would be useful to further improve the study implementing additional samples representative of the entire Italian population. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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14 pages, 1480 KiB

Open AccessArticle

Dental DNA as an Indicator of Post-Mortem Interval (PMI): A Pilot Research

by Ilenia Bianchi, Simone Grassi, Francesca Castiglione, Caterina Bartoli, Bianca De Saint Pierre, Martina Focardi, Antonio Oliva and Vilma Pinchi

Int. J. Mol. Sci. 2022, 23(21), 12896; https://doi.org/10.3390/ijms232112896 - 25 Oct 2022

Cited by 3 | Viewed by 1646

Abstract

Teeth have proven to be a reliable source of DNA for forensic analysis as the pulp is rich in cells and protected from damaging factors and contamination by dental hard tissues. The pilot study aims to evaluate the feasibility of Next-Generation sequencing analysis on dental pulp to detect genetic mutations in DNA caused by post-mortem cell necrosis. We used a 56-gene oncopanel kit on a sample of 17 teeth extracted from living patients. Time of the tooth avulsion was assumed as death of the individual and Post-mortem Interval (PMI) was the time elapse since the DNA extraction and analysis. Days and Accumulated Degree Days (ADD) were assumed as measures of PMI that ranged between 0 to 34 days. Only 38 of the 56 considered genes proved to be affected by mutations (101), thus being of forensic interest. More specifically, 14 mutations occurred only in a specific range of PMIs/ADD; 67 were detected (alone or as clusters of the same gene) at specific PMI/ADD; 22 occurred at every PMI/ADD, except for some specific intervals. Since dental pulp was not targeted by any oncological diseases and all teeth were intact, vital, and from patients with unremarkable medical history, it could be assumed that mutations were due to post-mortem DNA changes induced by pulp death and the increasing time elapse since death. This pilot study found encouraging results in the application of NGS analysis on dental DNA, especially for PMIs of several days for which the traditional tools for PMI estimation have limitations. Further research on a larger sample of PMI and validation research on a larger sample of PMI and validation of the results are indeed necessary. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

14 pages, 2124 KiB

Open AccessArticle

Do Circulating Redox Biomarkers Have Diagnostic Significance in Alcohol-Intoxicated People?

by Mateusz Maciejczyk, Iwona Ptaszyńska-Sarosiek, Anna Niemcunowicz-Janica, Michał Szeremeta, Napoleon Waszkiewicz, Agnieszka Kułak-Bejda, Urszula Cwalina, Miłosz Nesterowicz and Anna Zalewska

Int. J. Mol. Sci. 2022, 23(19), 11808; https://doi.org/10.3390/ijms231911808 - 05 Oct 2022

Cited by 2 | Viewed by 1303

Abstract

The toxic properties of ethanol are inextricably linked to oxidative stress. Despite many reports on the effects of alcohol dependence on blood redox homeostasis, there are no data on the oxidative stress profile in alcohol-poisoned cases. There are also no data on the diagnostic usefulness of redox biomarkers determined post-mortem in various biological fluids. This work investigates the utility of enzymatic and non-enzymatic antioxidant barrier, redox status, and oxidative/nitrosative stress biomarkers in different biological fluids (such as blood, urine, vitreous humor, and cerebrospinal fluid) in the post-mortem study of patients with acute alcohol intoxication. The study group included those who died due to acute ethanol intoxication (n = 22). The research showed a significant increase in glutathione peroxidase activity, total antioxidant status, ferric reducing antioxidant power, and tryptophan concentration only in the study group’s urine compared to the control. In other circulating fluids, both antioxidant enzyme activities and glycoxidation product concentrations were not significantly different in individuals who died of alcohol overdose compared with those who died suddenly. We also did not observe a connection between oxidation–reduction balance and the amount of alcohol consumed before death. These unexpected observations may be caused by irreversible post-mortem changes occurring at the cellular level due to autolysis and putrefaction. In summary, the use of circulating body fluids to assess redox homeostasis is limited in the post-mortem analysis. Our results indicate the increased stability of urine collected post mortem compared to other circulating bioliquids. Further studies are needed to assess the intensity of oxidative and carbonyl stress in ethanol-damaged organs and the effects of post-mortem processes on cellular redox balance. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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13 pages, 2676 KiB

Open AccessArticle

The Perfect Match: Assessment of Sample Collection Efficiency for Immunological and Molecular Findings in Different Types of Fabrics

by Sara C. Zapico, Alex Dytso, Leticia Rubio and Gabriela Roca

Int. J. Mol. Sci. 2022, 23(18), 10686; https://doi.org/10.3390/ijms231810686 - 14 Sep 2022

Cited by 4 | Viewed by 1516

Abstract

Body fluid identification at crime scenes can be crucial in retrieving the appropriate evidence that leads to the perpetrator and, in some cases, the victim. For this purpose, immunochromatographic tests are simple, fast and suitable for crime scenes. The potential sample is retrieved with a swab, normally a cotton swab, moistened in a specific buffer. Nonetheless, there are other swab types available, which have been proven to be efficient for DNA isolation and analysis. The aim of this study is to evaluate the efficiency of different swab types for body fluid identification as well as DNA isolation and characterization. Fifty microliters of human saliva were deposited in three different types of fabric (denim, cotton, and polyester). After 24 h at room temperature, samples were recovered by applying three different swab types, and the tests were performed. Subsequently, total DNA was recovered from the sample buffer. Cotton swabs performed worse in denim and cotton fabrics in both immunochromatography tests and DNA yield. No differences were observed for polyester. In contrast, and except for two replicates, it was possible to obtain a full DNA profile per fabric and swab type, and to identify the mtDNA haplogroup. In this paper, the impact of swab types on body fluid identification through the application of immunochromatographic tests is analyzed for the first time. This work corroborates previous research related to the influence of swab types in nuclear DNA isolation and characterization. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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Review

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20 pages, 672 KiB

Open AccessReview

The Revolution of Animal Genomics in Forensic Sciences

by Irene Cardinali, Domenico Tancredi and Hovirag Lancioni

Int. J. Mol. Sci. 2023, 24(10), 8821; https://doi.org/10.3390/ijms24108821 - 16 May 2023

Cited by 3 | Viewed by 2842

Abstract

Nowadays, the coexistence between humans and domestic animals (especially dogs and cats) has become a common scenario of daily life. Consequently, during a forensic investigation in civil or criminal cases, the biological material from a domestic animal could be considered “evidence” by law enforcement agencies. Animal genomics offers an important contribution in attacks and episodes of property destruction or in a crime scene where the non-human biological material is linked to the victim or perpetrator. However, only a few animal genetics laboratories in the world are able to carry out a valid forensic analysis, adhering to standards and guidelines that ensure the admissibility of data before a court of law. Today, forensic sciences focus on animal genetics considering all domestic species through the analysis of STRs (short tandem repeats) and autosomal and mitochondrial DNA SNPs (single nucleotide polymorphisms). However, the application of these molecular markers to wildlife seems to have gradually gained a strong relevance, aiming to tackle illegal traffic, avoid the loss of biodiversity, and protect endangered species. The development of third-generation sequencing technologies has glimmered new possibilities by bringing “the laboratory into the field”, with a reduction of both the enormous cost management of samples and the degradation of the biological material. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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19 pages, 1539 KiB

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Touch DNA Sampling Methods: Efficacy Evaluation and Systematic Review

by Pamela Tozzo, Enrico Mazzobel, Beatrice Marcante, Arianna Delicati and Luciana Caenazzo

Int. J. Mol. Sci. 2022, 23(24), 15541; https://doi.org/10.3390/ijms232415541 - 08 Dec 2022

Cited by 17 | Viewed by 3771

Abstract

Collection and interpretation of “touch DNA” from crime scenes represent crucial steps during criminal investigations, with clear consequences in courtrooms. Although the main aspects of this type of evidence have been extensively studied, some controversial issues remain. For instance, there is no conclusive evidence indicating which sampling method results in the highest rate of biological material recovery. Thus, this study aimed to describe the actual considerations on touch DNA and to compare three different sampling procedures, which were “single-swab”, “double-swab”, and “other methods” (i.e., cutting out, adhesive tape, FTA^® paper scraping), based on the experimental results published in the recent literature. The data analysis performed shows the higher efficiency of the single-swab method in DNA recovery in a wide variety of experimental settings. On the contrary, the double-swab technique and other methods do not seem to improve recovery rates. Despite the apparent discrepancy with previous research, these results underline certain limitations inherent to the sampling procedures investigated. The application of this information to forensic investigations and laboratories could improve operative standard procedures and enhance this almost fundamental investigative tool’s probative value. Full article

(This article belongs to the Special Issue Molecular Biology in Forensic Science: Past, Present and Future)

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