A Ge.F.I. - ISFG European collaborative study on DNA identification of Cannabis sativa samples using a 13-locus multiplex STR method.

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.

DNA mixtures interpretation - A proof-of-concept multi-software comparison highlighting different probabilistic methods' performances on challenging samples.

The present study investigated the capabilities and performances of semi-continuous and fully-continuous probabilistic approaches to DNA mixtures interpretation, particularly when dealing with Low-Template DNA mixtures. Five statistical interpretation software, such as Lab Retriever and LRmix Studio - involving semi-continuous algorithms - and DNA•VIEW®, EuroForMix and STRmixTM- employing fully-continuous formulae - were employed to calculate likelihood ratio, comparing the prosecution and the defense hypotheses relative to a series of on-purpose prepared DNA mixtures that respectively contained 2 and 3 known contributors. National Institute of Standards and Technologies (NIST) certified templates were used for samples set up, which contained different DNA amounts for each contributor. 2-person mixtures have been prepared with proportions equal to 1:1, 19:1 and 1:19 in terms of DNA concentration. Conversely, three person mixtures were constituted by proportions equal to 20:9:1, 8:1:1, 6:3:1 and 1:1:1 in terms of DNA concentration. Furthermore, 8 equally-proportioned 3-person mixtures were prepared by means of scalar dilutions starting from an overall amount of 0.500 ng, then ranging up to DNA samples with concentrations equal to 0.004 ng (i.e. Low-Template DNA). DNA mixtures were set up in triplicate and amplified with 7 DNA amplification kits (i.e. GlobalFiler PCR Amplification Kit, NGM SElect PCR Amplification Kit, MiniFiler PCR Amplification Kit, Power Plex Fusion, PowerPlex 6C Matrix System, Power Plex ESI 17 Fast and Power Plex ESX 17 Fast) in order to evaluate whether the selection of a certain kit might represent a bias factor, capable of altering the whole interpretation process. A multi-software approach helped us to highlight any trend in the likelihood ratio results provided by semi- and fully-continuous software. As a matter of fact, fully-continuous computations provided different (higher) results in terms of degrees of magnitude of the likelihood ratio values with respect to those from the semi-continuous approach, regardless of the amplification kit that was utilized.

Effect of different sports on body cell mass in highly trained athletes.

The objective of this study was to verify the impact of various sports on body cell mass (BCM). Ninety-eight male subjects, 17-33 years of age, participated in the study. The sample included athletes from three professional Italian football (soccer) teams, representing three different divisions (A, n=16; B, n=14; and C, n=18), judo athletes (J, n=10), and water polo athletes (W, n=14) who all competed at the national level. Twenty-six age-matched individuals served as the control group (CG). Fat-free mass (FFM), fat mass (FM), percent body fat (%BF), and BCM were assessed using bioimpedance spectroscopy (BIS). There were no significant differences in body weight and FFM among the groups. A and B were significantly taller than J and CG. B had a significantly lower body mass index (BMI; kg/m(2)) than CG, while C had a significantly lower BMI than J and CG. CG had a significantly greater FM and %BF than A, B, and C. C had a significantly lower BCM than Aand B. CG had a significantly lower BCM than A, B, J, and W. In conclusion, differences in BCM exist among athletes of different sports, and among athletes within the same sport who compete at different levels.

Analysis of body water compartments after a short sauna bath using bioelectric impedance analysis.

Studies have suggested that long-term sauna bathing may lower blood pressure in persons with hypertension by causing a direct loss of extracellular water and plasma minerals. The objective of the present study was to evaluate the effect of short-term sauna bathing on body water compartments as estimated by bioelectric impedance analysis (BIA). We recruited 15 men [mean age (+/-SD) of 23.93+/-5.12 years and mean body mass index (BMI) of 23.25+/-2.84 kg/m(2)] and 10 women matched for age and BMI. Total body resistance, reactance, and impedance were measured for all participants using BIA, at baseline, after a short sauna bath, and after a rest period. Total, extracellular, and intracellular water compartments were calculated using BIA formulae. There were no significant differences for any of the body water compartments when comparing the measurements taken before and after the sauna bath and after the rest period. However, it remains to be determined whether or not BIA is sensitive to rapid changes in water volume.