Forensic Pharmacogenetics

Current pharmacogenetics research in the clinical and medico-legal settings provides new options for disease treatment and prevention of ADR avoiding correlated death, and for screening interactions with the polymorphic P450 enzymes early on in drug development

Detection of AP-237 and synthetic cannabinoids on an infused letter sent to a German prisoner

In the past years, new psychoactive substances (NPS) started circulating in prisons, leading to health risks and challenges for the criminal justice system. Seizures of papers and cards impregnated with synthetic cannabinoid (SCs) have been reported. In November 2021, a letter suspected to be drug-infused was sent from a German prison to this laboratory. Toxicological analyses were performed by means of gas chromatography-mass spectrometry (GC-MS) for drug screening and liquid chromatography-tandem mass spectrometry (LC-MS/MS) as well as high-performance (HP) LC with diode-array detection (DAD) for semi-quantification of the compounds. The novel synthetic opioid (NSO) AP-237 was detected on the letter, with an estimated concentration of 1.2mug/cm2 , together with the SCs MDMB-4en-PINACA (77mug/cm2 ) and 5F-ADB (6.5mug/cm2 ). To the best of the authors' knowledge, this is the first time an NSO was detected on a drug-infused paper seized in a prison. Highly potent NSOs could easily be dissolved in organic solvents to produce impregnated papers and textiles, and this might represent a serious threat to the health of people in prison. Given the inhomogeneity in drug concentrations, health risks might in particular arise from the consumption of highly concentrated areas of the paper-so-called "hot spots"-especially when highly potent NSOs are used for infusion. Laboratories engaged in analyzing such impregnated papers should be aware of the potential presence of NSOs and adapt the respective methods accordingly

Ultrastructural and biochemical analysis of in vitro odontoblast differentiation

The purpose of this study was to develop an in vitro model of odontoblast differentiation and dentin formation which allows to deeply and better study the process of odontoblast differentiation and protein matrix deposition and mineralization. Human dental pulp cells were obtained from extracted molar teeth of donors under informed consent. Cells were induced to odontoblast differentiation for 7, 14, 21 and 28 days by adding in the cell medium dexamethasone, β-glicerophosphate, ascorbic acid and TGF-β1. Alizarin red staining, electron microscopy, western blotting were carried out to demonstrate the odontoblast differentiation process, extracellular matrix deposition and mineralization. Alizarin red staining results showed a gradual calcium deposition during the treatment, reaching the highest signal at 28 days of stimulation. Electron microscopy demonstrated the deposition of fibrillar structures in the extracellular matrix, connected with extracellular matrix proteins. Western blotting analysis showed the expression of collagen type I and dentin matrix protein 1, markers for the odontoblast phenotype. The in vitro odontoblast differentiation model described if an highly repetitive model and it represents a potential and promising tool to explore the extracellular matrix deposition and mineralization process in physiological and pathological conditions

Insights in opiates toxicity: impairment of human vascular mesenchymal stromal cells

: The most common pulmonary findings in opiate-related fatalities are congestion and oedema, as well as acute and/or chronic alveolar haemorrhage, the cause of which is thought to be a damage to the capillary endothelium related to ischemia. Human vascular mesenchymal stromal cells (vMSCs) play a fundamental role in tissue regeneration and repair after endothelial cell injury, and they express opioid receptors. The aim of this study was to assess the effect of in vitro morphine exposure on the physiological activity and maintenance of human vMSCs. vMSCs were obtained from abdominal aorta fragments collected during surgery repair and were exposed to incremental doses (0.1 mM, 0.4 mM, 0.8 mM and 1 mM) of morphine sulphate for 7 days. The effect was investigated through cell viability assessment, proliferation assay, reactive oxygen species (ROS) detection assay, senescence-associated β-galactosidase assay, senescent-related markers (p21WAF1/CIP1 and p16INK4) and the apoptosis-related marker caspase 3. Moreover, an ultrastructural analysis by transmission electron microscopy and in vitro vascular differentiation were evaluated. Results showed a decrease of the cellular metabolic activity, a pro-oxidant and pro-senescence effect, an increase in intracellular ROS and the activation of the apoptosis signalling, as well as ultrastructural modifications and impairment of vascular differentiation after morphine treatment of vMSC. Although confirmation studies are required on real fatal opiate intoxications, the approach based on morphological and immunofluorescence methodologies may have a high potential also as a useful tool or as a complementary method in forensic pathology. The application of these techniques in the future may lead to the identification of new markers and morphological parameters useful as complementary investigations for drug-related deaths

Morphological and immunoistochemical evaluation of cadaveric gingival specimens to estimate the post-mortem Interval

The estimation of the post-mortem interval (PMI) is a critical step in forensic medicine and remains one of the most challenging variables to determine. In general, the numerous methods currently used in estimating post-mortem interval yield to large post-mortem windows, are influenced by several factors and sometime contradict each other.  With the aim to obtain a much more accurate determination of the post-mortem interval we combined morphological ultrastructural and immunoistochemical analyses to reach a more detailed knowledge on tissue organization and degradation after death. Samples of gingival tissues obtained from 20 cadavers at different post-mortem intervals were processed for transmission electron microscopy to evaluate ultrastructural modifications in the epithelium and connective tissue. Gingival cells and the extracellular matrix of gingival tissues have been observed by a transmission electron microscopy (TEM) in combination with the evaluation of potential post-mortem biochemical markers, with the final goal to find a tight correlation between the ultrastructural modifications, the biomarker expression and the time of death. All the samples were also immunostained with anti-hypoxia-induced factor 1-α (HIF1-α) antibody, a transcription factor expressed in response to hypoxia, in order to evaluate the expression of HIF1-α, and to establish a correlation between the protein presence and the time of death. Results showed nuclear chromatin changes and cytoplasmic vacuolization in both epithelial and connective tissues and a different pattern of expression of HIF1alpha protein that correlate to the time of death. In conclusion, our preliminary findings suggest that ultrastructural investigations in combination with immunohistochemistry techniques in gingival specimens may represent a new tool to accurately and systematically estimate post-mortem interval

Come rileggere la sanità locale? Spagna e italia: esperienze a confronto. ¿Cómo impulsar la sanidad local? España e Italia: experiencia a debate

Il volume raccoglie le relazioni rivedute ed aggiornate svolte nell'ambito del convegno webinar "Come rileggere la Sanità locale? Spagna e Italia: esperienze a confronto" tenutosi l'11 marzo 2022, con il patrocinio della Scuola di Medicina e Chirurgia dell'Alma Mater Studiorum - Università di Bologna e della Real Academia de Medicina de Castilla-La Mancha, in collaborazione con CeSDirSan - Centro Interdisciplinare di Studi sul Diritto Sanitario. El volumen recoge las ponencias presentadas en el congreso webinar "¿Cómo impulsar la Sanidad Local? España e Italia: experiencia a debate" realizado el 11 de Marzo de 2022, bajo el patrocinio de la Facultad de Medicina de la Alma Mater Studiorum - Università di Bologna y de la Real Academia de Medicina de Castilla - La Mancha, con la colaboración de CeSDirSan - Centro Interdisciplinare di Studi sul Diritto Sanitario

A global analysis of Y-chromosomal haplotype diversity for 23 STR loci

In a worldwide collaborative effort, 19,630 Y-chromosomes were sampled from 129 different populations in 51 countries. These chromosomes were typed for 23 short-tandem repeat (STR) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385ab, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, GATAH4, DYS481, DYS533, DYS549, DYS570, DYS576, and DYS643) and using the PowerPlex Y23 System (PPY23, Promega Corporation, Madison, WI). Locus-specific allelic spectra of these markers were determined and a consistently high level of allelic diversity was observed. A considerable number of null, duplicate and off-ladder alleles were revealed. Standard single-locus and haplotype-based parameters were calculated and compared between subsets of Y-STR markers established for forensic casework. The PPY23 marker set provides substantially stronger discriminatory power than other available kits but at the same time reveals the same general patterns of population structure as other marker sets. A strong correlation was observed between the number of Y-STRs included in a marker set and some of the forensic parameters under study. Interestingly a weak but consistent trend toward smaller genetic distances resulting from larger numbers of markers became apparent.Peer reviewe

segreto professionale e d'ufficio

Tema ovog rada je judo kao dio paraolimpijskog sporta. Pretraživanjem domaće i strane literature te uvidom u rezultate istraživanja u radu su prikazani pozitivni uĉinci juda na funkcionalne sposobnosti (aerobna sposobnost i anaerobni energetski kapaciteti) i motoriĉke sposobnosti (kvantitativne-snaga, brzina, izdržljivost, fleksibilnost i kvalitativne-koordinacija, agilnost, ravnoteža, preciznost) te psihološke (samostalnost, samopouzdanje, motivacija) i socijalne osobine kod slijepih i slabovidnih osoba. U ovom diplomskom radu sažeta je povijest paraolimpijskog juda od samih zaĉetaka pa sve do danas. Opisane su vrijednosti juda kao i osnovne razlike treninga i judo borbi kod slijepih i slabovidnih judaša te su obraĊena pravila i klasifikacija slijepih i slabovidnih natjecatelja. U radu je proveden razgovor sa Tadijom Orlovićem koji je trenirao hrvatskog paraolimpijskog reprezentativca Mirsada Bećirovića i sa hrvatskom paraolimpijkom Lucijom Brešković. Kroz razgovore su iznjeli vrlo pouĉna zapažanja o ovom specifiĉnom naĉinu rada sa slijepim i slabovidnim osobama.Theme of this paper is judo as a part of Paralympic sport. Searching domestic and foreign literature and checking the results of researches, paper shows the positive effects of judo on functional ability (aerobic ability, anaerobic energetic capacity) and motor ability (quantitative- strength, speed, endurance, flexibility and qualitative-coordination, agility, balance, precision) as well as psychological (autonomy, self-confidence, motivation) and social features for blind and visually impaired persons. This thesis summarized the history of the Paralympic judo from the very beginning until today. It describes the values of judo and the basic differences of training and judo combat for blind and visually impaired judoka. In the paper are analysed judo rules, organization and classification of blind and visually impaired competitors. This paper contains interview with Tadija Orlović who trained Croatian Paralympic athlete Mirsad Bećirović and the Croatian paralympian Lucija Brešković. Through interviews they revealed very instructive observations on this particular methods of work with visually impaired people