Scopus Publications
- Consensus DNA profiling of formalin-fixed paraffin-embedded tissues: from genetic puzzle to a clear picture
Aldo DI NUNZIO, Fernanda IAFUSCO, Michele DI NUNZIO, Nicholas IACOVELLI, Roberto MIANO, Giulia SGAMBATI, Ciro DI NUNZIO, Nadia TINTO
Biochimica Clinica, 2025 - From dactyloscopy to the Minimum Surface Requirement (MSR): a metric to assess and obtain suitable human STR profiles
Michele Di Nunzio, Josep De Alcaraz-Fossoul, Vanessa Maroto-Nouveau, Clara Camprubí-Aumatell, Carme Barrot-Feixat
Forensic Science International, 2025
Friction skin ridge (Dactyloscopy) and DNA analyses have both played an essential role in forensic investigations for decades. The simultaneous and successful collection of a latent fingermark ridge pattern and the respective isolation of its human DNA can be a challenging task at a crime scene. Maximizing the quantity and quality of genetic information from this type of physical evidence has become one of the objectives of forensic science. Often, friction ridge impressions may lack discriminatory power due to low visual clarity of ridges and/or insufficient pattern area. In such cases, an optimized human DNA collection and extraction becomes crucial for obtaining a genetic identity from an unsuitable fingermark. Recent studies have proven that complete Short Tandem Repeat (STR) profiles can be acquired from touch DNA specimens. Although the impact of fingermark enhancement techniques on DNA recovery has been explored, no research has yet investigated the potential quantity of DNA that could be isolated per area of a ridge pattern. Here, DNA collection and extraction methods to determine the Minimum ridge Surface Requirement (MSR) to extract suitable human STR genotypes have been studied and optimized. Twelve participants, including biological males and females, provided ink prints of their index, middle, and ring fingers from both hands to calculate an average fingertip size to create a "standard" area for analysis across all donors. A standardized cutout template was created to fit every participant's latent fingermarks on pre-cleaned glass surfaces. Two fingermark "recharging-deposition-collecting" methods (i.e., Original Experiment and Optimized Experiment) and two swab types for DNA collection were compared (i.e., cotton swabs from Delta Lab (Spain) and 4N6FLOQSwabs® from Copan (Italy)). Then, ten participants deposited fingermarks for DNA extraction, quantification, STR amplification, and capillary electrophoresis (CE) for STR genotyping. Results showed that flocked swabs were more effective than cotton swabs at collecting DNA and produced suitable STR profiles with the optimized collection method. This research establishes the first MSR surfaces from which meaningful STR profiles can be acquired. These data provide key knowledge that assists crime investigators in prioritizing dactyloscopy, genotyping, or both analyses concurrently when examining latent fingermarks. - A Comparison of Convolutional Neural Networks using Transfer Learning for Cannabis Seed Classification
Ceur Workshop Proceedings, 2025 - Leveraging genetics to support forensic toxicology analysis: Demonstrating concordance among marijuana samples
Michele Di Nunzio, Maria Pieri, David Gangitano, Ciro Di Nunzio, Nadia Tinto, Massimo Niola, Carme Barrot-Feixat
Journal of Applied Research on Medicinal and Aromatic Plants, 2024
Cannabis sativa is an important plant for industrial purposes. Indeed, it is legal to cultivate and supply authorized low level Δ9-tetrahydrocannabinol (THC) cannabis plants for fiber and seeds (i.e., 0.2 % according to the European Union regulation on drugs). Today, chromatography/mass spectrometry-based procedures are successfully applied to determine THC concentration. Unfortunately, when cannabis samples have been degraded because wrongfully stored, toxicological results were affected, altering the final status of legal sample or illegal sample. Rather, genetic identification could give more information about the identity of these plants. For this reason, a degradation study was run to support how genetics can help to detect concordance in cannabis highly-degraded samples. Forty cannabis sub-samples were stored (for one week, two weeks, one month and two months) in auto-sealing bag to assess the effects of a wrong cannabis storage on weight reduction, genomic DNA changes, and corresponding STR profiles. Once respective time-period elapsed, each sub-sample’s weight was measured, the percentage weight loss calculated, and the genomic DNA was extracted and quantified, obtaining a complete STR profile for all samples. Furthermore, the aim of this study was to assess the same 13-loci short tandem repeat (STR) multiplex system used for the degradation experiment and evaluate the power of such markers in the identification and discrimination of highly degraded cannabis samples coming from real cases. The results of this study demonstrated that the 13-loci STR multiplex system successfully achieved the objective both for industry and forensic purposes. For each sample, all 13 loci were amplified, and degraded samples were correctly identified, suggesting that genetic typification could be a useful tool. The proposed procedure could be parallelly applied to toxicology analysis to detect if vegetable sample become from authorized plant, to help courts track back illegal samples or to achieve illegal cannabis genetic profiles for further comparison. - An initial exploration of machine learning for establishing associations between genetic markers and THC levels in Cannabis sativa samples
Selena Cisana, Michele Di Nunzio, Valentina Brenzini, Monica Omedei, Fabrizio Seganti, Christina Ververi, Enrico Gerace, Alberto Salomone, Andrea Berti, Filippo Barni, Sergio Schiavone, Andrea Coppi, Ciro Di Nunzio, Paolo Garofano, Eugenio Alladio
Forensic Science International Genetics, 2024 - How the forensic multidisciplinary approach can solve a fatal dog pack attack
M. Di Nunzio, A. Della Valle, A. Serino, F. Corrado, C. Di Nunzio
Forensic Science Medicine and Pathology, 2024
The authors present the case of a 61-year-old man found dead in an agricultural plot. The first investigation of the scene revealed the corpse laid face up in a spot of partially dried blood, next to an olive tree. His face, arms, legs, and abdomen showed signs of severe contusion and laceration of dogs’ bite wounds. Next to the victim, an olives bin had been found overturned on the ground. A multi-disciplinary approach, including crime scene analysis, autopsy findings, veterinary animals review, odontologist bite mark study, and forensic genetics DNA correlations, was performed. The present case is a documented watchdogs lethal pack attack and provides an example of how to recognize the more active participants thanks to their odontological alterations. It could be considered the first described dog pack attack case solved by dysgnathia alteration. Comparisons between the dental casts obtained from the dogs and the inflicted wounds were made, resulting in positive correlations between the injuries and the dental arches from two of the six involved dogs, thanks to dental abnormalities and DNA founding. The victim’s clothes were also compared with the dogs’ dental casts, confirming that they were the most active participants during the pack attack. Dogs’ DNA was finally matched with saliva traces found on victim’s clothes and skin bite marks. - Characterization and evaluation of nine Cannabis sativa chloroplast SNP markers for crop type determination and biogeographical origin on European samples
Michele Di Nunzio, Carme Barrot-Feixat, David Gangitano
Forensic Science International Genetics, 2024
Cannabis sativa can be classified in two main types, according to psychotropic cannabinoid ∆9-tetrahydrocannabinol (∆9-THC) content: the drug-type and the fiber-type. According to the European Monitoring Center for Drugs and Drug Addiction, most of the European Union countries consider the possession of cannabis, for personal use, a minor offense with possibility of incarceration. Despite of the model of legal supply (i.e., Spanish cannabis clubs, Netherlands coffee shops) or medical use (i.e., Italy), cannabis remains the most used and trafficked illicit plant in the European Union. Differentiating cannabis crops or tracing the biogeographical origin is crucial for law enforcement purposes. Chloroplast DNA (cpDNA) markers may assist to determine biogeographic origin and to differentiate hemp from marijuana. This research aims: to identify and to evaluate nine C. sativa cpDNA polymorphic SNP sites to differentiate crop type and to provide information about its biogeographical origin. Five SNaPshot™ assays for nine chloroplast markers were developed and conducted in marijuana samples seized in Chile, the USA-Mexico border and Spain, and hemp samples grown in Spain and in Italy. The SNapShot™ assays were tested on 122 cannabis samples, which included 16 blind samples, and were able to differentiate marijuana crop type from hemp crop type in all samples. Using phylogenetic analysis, genetic differences were observed between marijuana and hemp samples. Moreover, principal component analysis (PCA) supported the relationship among hemp samples, as well as for USA-Mexico border, Spanish, and Chilean marijuana samples. Genetic differences between groups based on the biogeographical origin and their crop type were observed. Increasing the number of genetic markers, including the most recently studied ones, and expanding the sample database will provide more accurate information about crop differentiation and biogeographical origin. - A customized protocol to generate STR profiles from latent fingerprints
Michele Di Nunzio, Ana María Rodríguez-Lozoya, Josep De Alcaraz-Fossoul, Carme Barrot-Feixat
Forensic Science International Genetics Supplement Series, 2022
For decades, dactyloscopical and DNA analyses have both played a key role in forensic investigations involving friction skin patterns and/or human biological material. In many occasions, friction ridge impressions may hold little discriminatory power due to low quality of ridge patterns and/or insufficient area extension of such patterns. In these cases, an appropriate human DNA collection and a high-quality DNA extraction become crucial steps to yield a genetic identity from an unsuitable latent fingerprint pattern. Indeed, over the past few years, it has been proven that complete Short Tandem Repeat (STR) profiles can be obtained from a touch DNA sample. In this study, a protocol has been customized to maximize the performance of genetic profiling from latent fingerprints. Six participants provided two sets of finger impressions on pre-cleaned glass surfaces. These impressions were generated by the participant’s dominant (DH) and non-dominant hand (NDH). Genetic material from fingerprints was pooled using a cotton swab for each donor and hand, combining 1–5 depositions consecutively. This was followed by DNA extraction, Real-Time PCR for DNA quantification, capillary electrophoresis (CE) for sequencing, and genotyping software for STR profiling. DNA yield was measured by ng/cm2 (DNA/fingerprint area). Statistical tests detected DNA yield differences by donor’s sex, age distribution, handedness and fingerprint pooling. Results revealed that DNA quantities from DH was dependent on the number of pooled fingerprints. However, NDH yielded similar DNA quantities across all fingerprint pooled combinations. With the aid of a customized protocol, DNA titers was improved and meaningful STR profiles were produced for donors’ DH and NDH. - Evaluation of genetic markers for the analysis of THC levels of Cannabis sativa samples using principal component analysis – A preliminary study
S. Cisana, M. Omedei, M. Di Nunzio, F. Seganti, V. Brenzini, A. Coppi, A. Berti, C. Di Nunzio, P. Garofano, E. Alladio
Forensic Science International Genetics Supplement Series, 2022 - A Ge.F.I. – ISFG European collaborative study on DNA identification of Cannabis sativa samples using a 13-locus multiplex STR method
M. Di Nunzio, V. Agostini, F. Alessandrini, C. Barrot-Feixat, A. Berti, C. Bini, M. Bottinelli, E. Carnevali, B. Corradini, M. Fabbri, P. Fattorini, P. Garofano, S. Gino, A. Mameli, A. Marino, C. Previderè, C. Robino, C. Romano, P. Tozzo, A. Verzeletti, L. Buscemi, D. Gangitano, C. Di Nunzio
Forensic Science International, 2021
Cannabis sativa is the most used controlled substance in Europe. With the advent of new and less restrictive European laws on cannabis sale for recreational use (including in Italy), an increase in indoor cannabis crops were observed. This increase was possible due to the availability of cannabis seeds through the internet market. Genetic identification of cannabis can link seizures and if in possession then might aid in an investigation. A 13-locus multiplex STR method was previously developed and validated by Houston et al. A collaborative exercise was organized by the Italian Forensic Geneticists - International Society of Forensic Genetics (Ge.F.I. - ISFG) Working Group with the aim to test the reproducibility, reliability and robustness of this multiplex cannabis STR kit. Twenty-one laboratories from three European countries participated in the collaborative exercise and were asked to perform STR typing of two cannabis samples. Cannabis DNA samples and the multiplex STR kit were provided by the University of Barcelona and Sam Houston State University. Different platforms for PCR amplification, capillary electrophoresis (CE) and genotyping software were selected at the discretion of the participating laboratories. Although the participating laboratories used different PCR equipment, CE platforms and genotyping software, concordant results were obtained from the majority of the samples. The overall genotyping success ratio was 96%. Only minor artifacts were observed. The mean peak height ratio was estimated to be 76.3% and 78.1% for sample 1 and sample 2, respectively. The lowest amount of -1 / + 1 stutter percentage produced, when the height of the parent allele was higher than 8000 RFU, resulted to be less than 10% of the parent allele height. Few common issues were observed such as a minor peak imbalance in some heterozygous loci, some artifact peaks and few instances of allelic drop-out. The results of this collaborative exercise demonstrated the robustness and applicability of the 13-locus system for cannabis DNA profiling for forensic purposes. - Bisphenol A impaired cell adhesion by altering the expression of adhesion and cytoskeleton proteins on human podocytes
Rafael Moreno-Gómez-Toledano, María I. Arenas, Clara González-Martínez, Nuria Olea-Herrero, Paula Reventún, Michele Di Nunzio, Sandra Sánchez-Esteban, Eduardo Arilla-Ferreiro, Marta Saura, Ricardo J. Bosch
Scientific Reports, 2020 - European validation of a Cannabis sativa 13-locus STR multiplex kit for genetic identification: A preliminary study
Michele Di Nunzio, Madeline Roman, Rachel Houston, Ciro Di Nunzio, David Gangitano, Carme Barrot-Feixat
Forensic Science International Genetics Supplement Series, 2019 - X-chromosome analysis in an unusual deficiency maternity case
Ciro Di Nunzio, Michele Di Nunzio, Flavio Saia, Pietrantonio Ricci, Aldo Di Nunzio, Silvano Presciuttini
Forensic Science International Genetics Supplement Series, 2019 - Analysis of recombination and mutation events for 12 X-Chr STR loci: A collaborative family study of the Italian Speaking Working Group Ge.F.I
C. Bini, C. Di Nunzio, S. Aneli, S. Sarno, M. Alù, E. Carnevali, E. Colao, M. Di Nunzio, M. Fabbri, P. Fattorini, P. Grignani, A. Piccinini, E. Ponzano, C. Robino, A. Rocchi, F. Scarnicci, C. Turchi, A. Verzeletti, S. Pelotti
Forensic Science International Genetics Supplement Series, 2019 - A GEFI collaborative exercise on DNA/RNA co-analysis and mRNA profiling interpretation
E. Carnevali, D. Lacerenza, S. Severini, F. Alessandrini, C. Bini, C. Di Nunzio, M. Di Nunzio, M. Fabbri, P. Fattorini, A. Piccinini, E. Ponzano, G. Portera, C. Previderè, A. Renieri, F. Scarnicci, A. Verzeletti, S. Pelotti, M. van den Berge, T. Sijen, C. Robino
Forensic Science International Genetics Supplement Series, 2017
