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. (c) 2021 Elsevier B.V. All rights reserved.

Estimation of the time since death is a practical task in daily forensic casework but available methods lack reliability especially in complex deaths and after long PM period. MirRNA analysis should be ideally an useful ancillary tool as it proved to be sensitive in forensics especially for body fluid identification.Here we analyzed 10 miRNAs with a supposed role in circadian rhythms through an RT-qPCR assay in postmortem samples of blood (n = 12) and vitreous humor (n = 12) from individuals died in the day or at night, in order to find those with an oscillating pattern of variation. The expression stability of four endogenous controls was also tested to find the most suitable for normalization. SNORD95 proved to be the best and was used in both body fluids. Four miRNAs showed significant differential expression between individuals died at daytime and at nighttime, mir-106b and mir-96 in vitreal samples and mir-142-5p and mir-219 in blood. Results are preliminary and limited to the small sample set. Future studies on more samples and with additional markers are needed to further elucidate the role of miRNA profiling in postmortem contexts and how useful they would be as "chronobiomarkers" for time of death determination. (C) 2015 Published by Elsevier Ireland Ltd.

The ambitious idea of using a short piece of DNA for large-scale species identification (DNA barcoding) is already a powerful tool for scientists and the application of this standard technique seems promising in a range of fields including forensic genetics. While DNA barcoding enjoyed a remarkable success for animal identification through cytochrome c oxidase I (COI) analysis, the attempts to identify a single barcode for plants remained a vain hope for a longtime. From the beginning, the Consortium for the Barcode of Life (CBOL) showed a lack of agreement on a core plant barcode, reflecting the diversity of viewpoints. Different research groups advocated various markers with divergent set of criteria until the recent publication by the CBOL-Plant Working Group. After a four-year effort, in 2009 the International Team concluded to agree on standard markers promoting a multilocus solution (rbcL and matK), with 70-75% of discrimination to the species level. In 2009 our group firstly proposed the broad application of DNA barcoding principles as a tool for identification of trace botanical evidence through the analysis of two chloroplast loci (trnH-psbA and trnL-trnF) in plant species belonging to local flora. Difficulties and drawbacks that were encountered included a poor coverage of species in specific databases and the lack of authenticated reference sequences for the selected markers. Successful preliminary results were obtained providing an approach to progressively identify unknown plant specimens to a given taxonomic rank, usable by any non-specialist botanist or in case of a shortage of taxonomic expertise. Now we considered mandatory to update and to compare our previous findings with the new selected plastid markers (matK+rbcL), taking into account forensic requirements. Features of all the four loci (the two previously analyzed trnH-psbA+trnL-trnF and matK+rbcL) were compared singly and in multilocus solutions to assess the most suitable combination for forensic botany. Based on obtained results, we recommend the adoption of a two-locus combination with rbcL+trnH-psbA plastid markers, which currently best satisfies forensic needs for botanical species identification.

Large-scale population studies have proved that genetic factors contribute to individual differences in smoking behavior. Genes responsible for nicotine's pharmacokinetics and pharmacodynamics seem mainly involved, although a significant fraction of variance remains unexplained. In this study we examined 10 SNPs from 8 candidate genes with positive previous reports of association with smoking. A total of 454 Italian unrelated subjects were genotyped by a multiplex minisequencing assay through the SNaPShot kit. Cases were chosen as current and former nicotine dependent (FTND≥4 and SQ≥15), while controls were smoking-exposed but non-dependent and never smoker individuals (FTND=0 and SQ≤10 and FTND=0 and SQ=0, respectively). Preliminary results shows that the SNPs CHRNA5-rs16969968 and CHRNA3-rs1051730 could be associated with risk of developing nicotine dependence. Factors as age, sex, and exposition to smoke were also found as possible factors of risk of nicotine addiction. The identification of susceptibility loci for individual response to substance abuse is particularly motivating for medicine for the global epidemic dimension of addictions and the urgent need of effective preventive and therapeutic strategies.

Suicide is a devastating psychopathological trait with familial transmission. According to epidemiological data suicide is partly under genetic influence. Despite the extensive body of association studies, to date genetic factors linked to suicide diathesis are largely unknown. In this study a set of 8 SNPs and a 43bp ins/del was genotyped in an Italian sample of suicide completers (n=65) and control subjects (n=77). The selected markers are spread across multiple brain pathways: serotonergic, stress-related HPA axis, catecholamine metabolism and nitric oxide (NO) neurotransmission. Basic analysis at single loci shows significant differences in allele and genotype distributions between victims and controls for TPH1-rs1800532(allele A) and NOSI-rs693534(allele A), pointing to an association with the group of suicides. Presented results are intended to be exploratory, requiring further investigation in a larger sample size. Systematic efforts in suicide research is quite important in order to empower understanding of risk factors and developing preventive strategies in clinical practice.

Substance addiction is a complex chronic brain disorder, characterized by neurobiological changes leading to compulsive drug seeking and taking. Although environmental factors contribute to drug addiction, evidence showed that genetic factors with multiple genes also play a significant role. Cannabis and tobacco result as the most common widely abused substances. Epidemiological studies have strongly implicated genetics in nicotine and marijuana consuming and vulnerability to subsequent dependence, estimating the range of heritability from 34% to 78% for cannabis and approximately from 50% to 70% for nicotine. Furthermore, varying aspects of impulsive personality and principal psychiatric disorders co-occur with tobacco and cannabis dependence status. We evaluate the possibility of identifying an individual's risk probability to become an addict, based on a genotype analysis and the different possible applications in legal medicine and forensic genetics.

Botanical evidence can provide useful leads in forensic investigations, however many plant materials cannot be identified to the species level only by morphological features. Taking advantage of DNA sequencing and other biomolecular techniques, exact identification of plants becomes useful in selected cases in court producing successful results. We propose a new barcoding markers for plant molecular systematic investigations at the species level by universal primers amplifications.