Human DNA contamination of postmortem examination facilities: Impact of COVID-19 cleaning procedure

The DNA contamination of evidentiary trace samples, included those collected in the autopsy room, has significant detrimental consequences for forensic genetics investigation. After the COVID-19 pandemic, methods to prevent environmental contamination in the autopsy room have been developed and intensified. This study aimed to evaluate the level of human DNA contamination of a postmortem examination facility before and after the introduction of COVID-19-related disinfection and cleaning procedures. Ninety-one swabs were collected from the surfaces and the dissecting instruments, analyzed by real-time quantitative PCR (q-PCR), and typed for 21 autosomal STRs. Sixty-seven out of 91 samples resulted in quantifiable human DNA, ranging from 1 pg/μl to 12.4 ng/μl, including all the samples collected before the implementation of COVID-19 cleaning procedures (n = 38) and 29 out of 53 (54.7%) samples taken afterward. All samples containing human DNA were amplified, resulting in mixed (83.6%), single (13.4%), and incomplete (3%) profiles. A statistically significant decrease in DNA contamination was found for dissecting instruments after treatment with chlorhexidine and autoclave (p< 0.05). Environmental decontamination strategies adopted during the COVID-19 pandemic only partially solved the long-standing issue of DNA contamination of postmortem examination facilities. The pandemic represents an opportunity to further stress the need for standardized evidence-based protocols targeted to overcome the problem of DNA contamination in the autopsy room

Development and validation of a rapid LC-MS/MS method for the detection of 182 novel psychoactive substances in whole blood

INTRODUCTION: The analysis of novel psychoactive substances (NPS) represents a challenge in forensic toxicology, due to the high number of compounds characterized by different structures and physicochemical properties both among different subclasses and within a single subclass of NPS. The aim of the present work is the development and validation of a targeted liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the detection of NPS in whole blood.MATERIALS AND METHODS: A protein-precipitation based LC-MS/MS method for the detection of more than 180 NPS was developed and validated by assessing the following parameters: selectivity, linearity, accuracy, precision, limit of detection (LOD) and of quantification (LOQ) recovery, and matrix effect. Then, the method was applied to real forensic samples.RESULTS: The method allowed the identification of 132 synthetic cannabinoids, 22 synthetic opioids, and 28 substances among synthetic cathinones, stimulants, and other drugs. Validation was successfully achieved for most of the compounds. Linearity was in the range of 0.25-10ng/ml for synthetic cannabinoids and 0.25-25ng/ml for other drugs. Accuracy and precision were acceptable according to international guidelines. Three cases tested positive for fentanyl and ketamine, in the setting of emergency room administration.CONCLUSIONS: The present methodology represents a fast, not expensive, wide-panel method for the analysis of more than 180 NPS by LC-MS/MS, which can be profitably applied both in a clinical context and in postmortem toxicology

Infection Induced Fetal Inflammatory Response Syndrome (FIRS): State-of- the-Art and Medico-Legal Implications—A Narrative Review

Fetal inflammatory response syndrome (FIRS) represents the fetal inflammatory reaction to intrauterine infection or injury, potentially leading to multiorgan impairment, neonatal mortality, and morbidity. Infections induce FIRS after chorioamnionitis (CA), defined as acute maternal inflammatory response to amniotic fluid infection, acute funisitis and chorionic vasculitis. FIRS involves many molecules, i.e., cytokines and/or chemokines, able to directly or indirectly damage fetal organs. Therefore, due to FIRS being a condition with a complex etiopathogenesis and multiple organ dysfunction, especially brain injury, medical liability is frequently claimed. In medical malpractice, reconstruction of the pathological pathways is paramount. However, in cases of FIRS, ideal medical conduct is hard to delineate, due to uncertainty in diagnosis, treatment, and prognosis of this highly complex condition. This narrative review revises the current knowledge of FIRS caused by infections, maternal and neonatal diagnosis and treatments, the main consequences of the disease and their prognoses, and discusses the medico-legal implications

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

L'effetto psicoattivo della "Cannabis Light". Dalla legge italiana alla psicofarmacologia

In 2019, the Italian Supreme Court established that hemp, for non-medical use, cannot be commercialized for human use, when the “psychotropic effect” of the product or its “offensiveness” can be demonstrated. The first chapter of this work reports a review of the European and Italian legislation on hemp cultivation, as well as the hemp production chain and commercial activities. The second chapter reports the pharmacological aspects and the psychoactive effects of light cannabis, along with pharmacokinetics of the main Cannabis compounds: Δ9-tetrahydrocannabinol (Δ9-THC), Cannabidiol (CBD) and Cannabinol (CBN). The aim of the experimental study, reported in the third chapter, is to assess Δ9-THC and CBD blood concentrations after smoking “light cannabis”, and its effects on vigilance, cognitive and motor skills. Eighteen young adults consumed three light cannabis cigarettes with a percentage of 0.41% of Δ9-THC and of 12.41% of CBD. Blood samples were collected before the experiment (t0) and after pre-defined time-lapses. Five performance tasks and a subjective scale were employed for measuring cognitive and psychomotor performances the day before the experiment (TT0) and after the third cigarette (TT1). Mean (SD) concentrations (ng/ml) were between 1.0(0.8) in t1 and 0.3(0.3) in t5 for Δ9-THC; and 10.5(10.3) in t1 and 5.7(5.7) in t5 for CBD. No significant differences were observed between TT0 and TT1 for all performed psychomotor performance task. Δ9-THC and CBD concentrations showed a high inter-subject variability, and the average concentrations were lower than those previously reported. Toxicological results showed a decrease of Δ9-THC and CBD after the third light cannabis cigarette, and a Δ9-THC /CBD ratio always < 1 was observed. This value might be useful in discriminating light cannabis versus illegal/medical cannabis consumption. The lack of impairment observed in our participants can be interpreted as a consequence of the very low concentrations in the blood.Nel 2019, una sentenza della Suprema Corte di Cassazione ha stabilito che la canapa, al di fuori degli usi medici, non può essere commercializzata se destinata al consumo umano, qualora sia dimostrata la "capacità drogante" del prodotto. Il primo capitolo dell’elaborato di tesi riporta una revisione della legislazione Italiana ed Europea sulla coltivazione della canapa e sulla catena produttiva e di commercio. Il secondo capitolo presenta gli aspetti farmacologici dei principali composti della canapa, il Δ9-tetraidrocannabinolo(Δ9-THC), il Cannabidiolo(CBD) ed il Cannabinolo(CBN). Il terzo capitolo riporta lo studio sperimentale, il cui scopo è quello di studiare la farmacocinetica e le proprietà psicotrope della cosiddetta "cannabis light". Diciotto giovani adulti hanno consumato in condizioni sperimentali tre sigarette contenenti cannabis light, con una percentuale di Δ9-THC dello 0.41% e di CBD del 12.41%. Sono stati raccolti ed analizzati campioni ematici prima dell'esperimento (t0) e dopo periodi di tempo predefiniti. Cinque test di performance psicomotori sono stati somministrati il giorno prima dell'esperimento (TT0) e dopo la terza sigaretta (TT1). Le concentrazioni medie (Dev.st.) in ng/ml rilevate erano comprese tra 1.0(0.8) in t1 e 0.3(0.3) in t5 per il Δ9-THC; e tra 10.5(10.3) in t1 e 5.7(5.7) in t5 per il CBD. Non sono state osservate differenze significative tra i risultati dei test psicomotori tra TT0 e TT1. Il Δ9-THC ed il CBD hanno mostrato un'ampia variabilità inter-individuale, e le concentrazioni medie osservate nel presente studio si sono rilevate inferiori a quelle già riportate su popolazioni ridotte. E’ stata osservata una riduzione della concentrazione di entrambi i composti dopo la terza sigaretta, ed un rapporto Δ9-THC/CBD sempre inferiore a 1. Questo valore potrebbe essere utilizzato per discriminare tra utilizzo di cannabis light e cannabis con elevata percentuale di Δ9-THC. La mancanza di impairment psicomotorio potrebbe essere interpretata come una conseguenza delle basse concentrazioni ematiche

PANDEMIA DA COVID-19 E ABUSO DI SOSTANZE STUPEFACENTI. LE NUOVE FRONTIERE DELLA RIDUZIONE DEL DANNO

Il modello di riduzione del danno (RdD), una misura sanitaria già diffusa in Europa dall’inizio degli anni ’80, ha come principale obiettivo quello di tutelare la salute dell’individuo che fa uso di sostanze stupefacenti. La diffusione del COVID-19 ha creato nuove aree di vulnerabilità ed esacerbato quelle preesistenti, ponendo sfide senza precedenti in ambito sanitario, tra cui la tutela delle persone che consumano droghe. Per le persone che utilizzano sostanze stupefacenti sono state diffuse buone pratiche, che si aggiungono alle raccomandazioni standard. Non di meno, le attuali politiche di RdD dovranno essere potenziate alla luce della nuova pandemia COVID-19, come previsto in questi mesi dalle principali società scientifiche ed associazioni nazionali ed internazionali. I messaggi sulla RdD incentrati sui rischi associati all’iniezione dovranno essere rimodulati alla luce del contagio attraverso droplets, tipico di pratiche considerate fino ad oggi “meno a rischio”, come la condivisione di sigarette, strumenti per la vaporizzazione e l’inalazione. L’assistenza territoriale dovrà essere potenziata con l’ausilio su larga scala della telemedicina, percorso già avviato nei primi due mesi della pandemia, nonché per mezzo di screening tossicologici ed infettivologici a domicilio, sulla base del modello applicato nella provincia di Hubei (Cina). L’attività dei laboratori di tossicologia forense e clinica e l’attivazione di ulteriori sistemi di allerta rapida saranno indirizzati alla prevenzione e alla tempestiva segnalazione delle possibili variazioni del mercato in termini di modifica della domanda, sostituzione e contraffazione delle sostanz

Misinterpretation of Anogenital Findings and Misdiagnosis of Child Sexual Abuse: The Role of the Forensic Pathologist

Background: The interpretation of anogenital postmortem findings is an issue of main concern, because the nature and appearance of anogenital tissues during the postmortem interval is not widely known by health providers. Case: An 8-year-old girl died in the hospital 48 hours after hospitalization. On the basis of the atypical anogenital findings, the health care professionals notified the fact to the Public Prosecutor as an alleged child abuse. The forensic pathologist ruled out this possibility, interpreting the anal findings due to physiological postmortem anal alterations and to the insertion of suppositories before death. Summary and Conclusion: Forensic pathological analysis should be included in routine postmortem evaluation in the case of suspected child sexual abuse, because normal postmortem findings could be misinterpreted by physicians, whose sole experience is on the basis of antemortem scenarios

Sequential development of hepatocellular carcinoma and liver angiosarcoma in a vinyl chloride\u2013exposed worker

Strong experimental and clinical evidences have definitely linked occupational vinyl chloride exposure to development of angiosarcoma of the liver. In contrast, despite the International Agency for Research on Cancer having included vinyl chloride among the causes of hepatocellular carcinoma, the association between vinyl chloride exposure and hepatocellular carcinoma remains debated. This issue is relevant, because occupational exposure to high levels of vinyl chloride may still occur. We report a unique case of sequential occurrences of hepatocellular carcinoma and angiosarcoma of the liver, in a vinyl chlorideâ\u80\u93exposed worker without cirrhosis and any known risk factor for chronic liver disease. Both the hepatocellular carcinoma and the surrounding normal liver showed micronucleus formation, which reflects genotoxic effect of vinyl chloride. Angiosarcoma showed a KRAS G12D point mutation, which is considered to be characteristic of vinyl chlorideâ\u80\u93induced angiosarcoma. This case supports the pathogenic role of vinyl chloride in both hepatocellular carcinoma and angiosarcoma development

Methodology for the identification of vulnerable asylum seekers

Asylum seekers often experience situations of vulnerability, being frequently exposed to a heightened risk of harm, and thus require special care, support and protection. The categories of â\u80\u9cvulnerable personsâ\u80\u9d, identified by International Legislation, and an individualâ\u80\u99s classification as a â\u80\u9cvulnerable asylum seekerâ\u80\u9d, have important implications in the reception procedures, in the decision-making phase and in the definition of therapeutic needs and rehabilitation. The Istanbul Protocol, the first international guideline approved by the United Nations and applied in different contexts, is not applicable for the assessment of the totality of the conditions (medical and otherwise), and therefore, the identification and assessment of conditions of vulnerability is largely delegated to questionnaires administered by non-medical personnel. The proposed methodology, based on the modificatory reworking of the Guidelines of the International Academy of Legal Medicine concerning the â\u80\u9cmedicolegal ascertainment of personal injury and damage on the living personâ\u80\u9d, takes into consideration all the medical issues relevant for the decision concerning the applicant, both in the reception procedures and in the outcome of the asylum application