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The sensitivity and specificity of a novel method of screening for cocaine in hair, based on matrix-assisted laser desorption/ionisation (MALDI) mass spectrometry (MS), have been evaluated. The method entails a rapid extraction procedure consisting of shaking 2.5 mg pulverised hair at high frequency in the presence of an acidic solution (160 µL of water, 20 µL of acetonitrile and 20 µL of 1 M trifluoroacetic acid) and a stainless-steel bullet. Following centrifugation, the supernatant is dried under a nitrogen stream, and the residue is reconstituted in 10 µL of methanol/trifluoroacetic acid (7:3; v/v). One microlitre of the extract is deposed on a MALDI sample holder previously scrubbed with graphite; an α-cyano-4-hydroxycinnamic acid (matrix) solution is electrosprayed over the dried sample surface to achieve a uniform distribution of matrix crystals. The identification of cocaine is obtained by post-source decay experiments performed on its MH⁺ ion (m/z 304), with a limit of detection of 0.1 ng/mg of cocaine. A total of 304 hair samples were analysed in parallel by MALDI-MS and a reference gas chromatography-MS method. The obtained results demonstrate specificity and sensitivity of 100% for MALDI-MS. Evidence of cocaine presence was easily obtained even when hair samples exhibiting particularly low cocaine levels (<0.5 ng/mg) were analysed.

Estimation of the firing range is often critical for reconstructing gunshot fatalities, where the main measurable evidence is the gunshot residue (GSR). In the present study intermediate-range gunshot wounds have been analysed by means of a micro-computed tomography (micro-CT) coupled to an image analysis software in order to quantify the powder particles and to determine the firing distance. A total of 50 shootings were performed on skin sections obtained from human legs surgically amputated for medical reasons. For each tested distance (5, 15, 23, 30 and 40 cm), firing was carried out perpendicularly at the samples using a 7.65-mm pistol loaded with jacketed bullets. Uninjured skin sections were used as controls. By increasing the firing distance, micro-CT analysis demonstrated a clear decreasing trend in the mean GSR percentage, particularly for shots fired from more than 15 cm. For distances under 23 cm, the powder particles were concentrated on the epidermis and dermis around the hole, and inside the cavity; while, at greater distances, they were deposited only on the skin surface. Statistical analysis showed a nonlinear relationship between the amount of GSR deposits and the firing range, well explained by a Gaussian-like function. The proposed method allowed a good discrimination for all the tested distances, proving to be an objective, rapid and inexpensive tool for estimating the firing range in intermediate-range gunshot wounds.

Advances in personalized medicine and Systems Biology have introduced probabilistic models and error discovery to cardiovascular care, aiding disease prevention and procedural planning. However, clinical application faces cultural, technical, and methodological hurdles. Patient autonomy remains essential, with shared decision-making (SDM) gaining importance in managing complex cardiovascular treatment options. Effective SDM relies on collaboration between providers and patients, guided by P5 Medicine principles, which combine psycho-cognitive considerations with predictive, personalized, preventive, and participatory care. Here we propose a 3-step methodological proposal for implementing SDM and enhancing consent acquisition in cardiovascular care. The approach emphasizes personalized patient engagement and the need for clear, comprehensive consent processes. It identifies and addresses significant gaps in current practices, including the complexity of consent language, information dispersion, and the specific needs of vulnerable populations. Issues of Medical Responsibility and/or Liability may raise in the case of absence of consent acquisition or invalid consent due to insufficient/incorrect information. The International Guidelines on Medico-Legal Methods of Ascertainment and Evaluation Criteria are reported. In conclusion, the paper proposes practical solutions, including the use of artificial intelligence (AI) to enhance decision-making and patient counseling, and strategies to ensure that consent processes are both thorough and legally sound and respectful to the individual's autonomy.

Anabolic androgenic steroids (AAS) are the main class of doping agents and their consumption produces adverse effects involving several organs and systems. Three cases of sudden cardiac death (SCD) and one of death due to congestive heart failure of previously healthy athletes who were AAS users are herein reported. Concentric cardiac hypertrophy with focal fibrosis (one case), dilated cardiomyopathy with patchy myocyte death (two cases) and eosinophilic myocarditis (one case) were observed and most probably relate to the final event. Molecular investigation for viral genomes was positive in one case (Ebstein virus). Our data confirm previous findings, showing that the most typical cardiac abnormality in AAS abusers is left ventricular hypertrophy, associated with fibrosis and myocytolysis. An exceptional cardiovascular substrate was represented by the case with drug induced eosinophilic myocarditis. These features are at risk of ventricular arrhythmias as well as congestive heart failure. The cause–effect relationship between AAS abuse and cardiac death can be established only by a rigorous methodology with the use of standardized protocols, including precise morphological studies of all target organs to search for chronic toxic effects. Laboratory investigations should focus on AAS searching on a wide range of biological matrices to demonstrate type, magnitude and time of exposure.

Embalming and formalin fixation are common, and yet they can create problems for the forensic scientist if a drug has been the cause of death and if the only available specimens to be analyzed are formalin-fixed tissues. Previous studies have demonstrated that during fixation xenobiotics are extracted into formalin according to tissue and fixing solution characteristics. In some cases formalin can react with the analyte resulting in the production of new chemical entities. Regarding cocaine and its metabolites, Cingolani et al. have reported that formalin-fixation extracts benzoylecgonine (BE) from tissues and that BE is stable in the fixing solution. However, the stability and kinetic properties of cocaine remain so far unexplored. Our data show that in buffered formalin (pH 7.4) cocaine is hydrolyzed to BE in agreement with a pseudo first-order reaction kinetic (half-life time ∼7 days), whereas in unbuffered formalin (pH ∼ 3.5) it is relatively stable over a period of 30 days. The analysis of brain and liver samples at different fixation times indicates that during fixation an extraction process occurs for both analytes and that the extraction is more efficient in the liver than in the brain, probably because of a greater lipophilicity of the brain tissue. In conclusion, our study demonstrates that formalin-fixed tissues and their fixing solutions can be used for cocaine analysis only if a short time period has passed since the fixation beginning. The rapid extraction process of cocaine into formalin and the concomitant hydrolysis to BE occurring in buffered formalin may prevent the identification of cocaine in both tissues and formalin solution already at 15–30 days after fixation. Moreover, the unpredictable extraction rate of both analytes, along with the hydrolysis of cocaine into BE significantly affects tissue concentrations, thus complicating the interpretation of quantitative results.

We describe a new method, based on indirect measures of implicit autobiographical memory, that allows evaluation of which of two contrasting autobiographical events (e.g., crimes) is true for a given individual. Participants were requested to classify sentences describing possible autobiographical events by pressing one of two response keys. Responses were faster when sentences related to truly autobiographical events shared the same response key with other sentences reporting true events and slower when sentences related to truly autobiographical events shared the same response key with sentences reporting false events. This method has possible application in forensic settings and as a lie-detection technique.

Part 1 of the review “ Back to the Future ” examines the historical evolution of the medico-legal autopsy and microscopy techniques, from Ancient Civilization to the Post-Genomic Era. In the section focusing on “ The Past ”, the study of historical sources concerning the origins and development of the medico-legal autopsy, from the Bronze Age until the Middle Ages, shows how, as early as 2000 BC, the performance of autopsies for medico-legal purposes was a known and widespread practice in some ancient civilizations in Egypt, the Far East and later in Europe. In the section focusing on “ The Present ”, the improvement of autopsy techniques by Friedrich Albert Zenker and Rudolf Virchow and the contemporary development of optical microscopy techniques for forensic purposes during the 19th and 20th centuries are reported, emphasizing, the regulation of medico-legal autopsies in diverse nations around the world and the publication of international guidelines or best practices elaborated by International Scientific Societies. Finally, in “The Future” section, innovative robotized and advanced microscopy systems and techniques, including their possible use in the bio-medicolegal field, are reported, which should lead to the improvement and standardization of the autopsy methodology, thereby achieving a more precise identification of natural and traumatic pathologies.

Ketamine (KT), primarily used as a general anaesthetic agent in clinical practice, has become very popular in recent years as a recreational drug, due to its dissociative and hallucinogenic effects. A liquid chromatography-high resolution mass spectrometry (LC–HRMS) method has been developed and validated for the quantification of KT and its main metabolite norketamine (NK) in 2.0mg of hair. Sample preparation consisted of a rapid, simultaneous pulverization and extraction step in acidic solution, followed by centrifugation and filtration. Gradient elution was performed by an Atlantis T3 analytical column, and deuterated KT was used as the internal standard. Positive ion electrospray ionization and HRMS determination in full-scan mode were achieved with an Orbitrap mass spectrometer. The method has a linear range of 0.05–50ng/mg, a limit of quantisation of 0.05ng/mg and a limit of detection of 0.02ng/mg for both KT and NK. The validated method was applied for the determination of KT and NK in two authentic hair samples from subjects suspected of taking psychoactive substances. The detection of the metabolite at low concentration gave proof for systemic drug origin and an investigation into the possible presence of further metabolites was performed by means of retrospective screening.