Julia Lazzari

Background/Objectives: 3,4-Methylenedioxy-alpha-pyrrolidinohexanophenone (MDPHP) is a pyrrolidinophenone synthetic cathinone whose post-mortem interpretation is particularly difficult when advanced decomposition prevents blood-based toxicology. This case report examined instrument-derived semiquantitative MDPHP values in solid tissues and necrophagous larvae and assessed the complementary evidential value of larval matrices. Methods: Post-mortem computed tomography, complete autopsy, histology, and liquid chromatography–high-resolution mass spectrometry (LC-HRMS) were performed in an adult decedent found in advanced decomposition. Brain, liver, kidney, and four separately collected larval groups were analyzed using a procedure previously validated in blood but not validated in tissues or larvae. Results: No fatal traumatic lesion was identified. Pulmonary edema and myocardial fibrofatty replacement were documented. Instrument-derived semiquantitative MDPHP values were 0.45 ng/mg in brain, 0.75 ng/mg in liver, and 2.44 ng/mg in kidney; the four larval groups yielded <0.06, 0.43, 1.58, and 0.22 ng/mg, respectively. These values are exploratory and semiquantitative because matrix-specific validation data are unavailable. The Larvae III extract also showed signals tentatively assigned to M3.1, M5.2, and M9.1. Conclusions: Necrophagous larvae preserved evidence of MDPHP exposure and phase-I metabolite signals. The findings do not establish validated quantitative concentrations, a lethal threshold, or a tissue-to-larva relationship. In the integrated medicolegal context, MDPHP intoxication remained a plausible contributor, but causal interpretation was limited by the single-case design, decomposition, underlying myocardial pathology, absence of conventional matrices, and lack of tissue- and larva-specific method validation.

Estimating the post-mortem interval (PMI) using microRNAs (miRNAs) in vitreous humor (VH) is a promising technique in forensic pathology. However, the reliability of quantitative Real-Time PCR (qPCR) data in this matrix is currently constrained by a critical methodological challenge: the lack of a rigorously validated endogenous reference gene (normalizer) capable of correcting for non-biological variations without being influenced by decomposition. This study aimed to identify a robust reference gene for VH analysis by performing a comparative validation of two candidates proposed in the literature: miR-222-3p and miR-96-5p. VH samples were collected from 47 forensic autopsy cases with estimated PMIs ranging from 3 to 24 h. The validation process assessed three key parameters: amplification detectability, expression stability (Coefficient of Variation, CV), and statistical independence from both the PMI and the pre-analytical freezing interval using regression models. MiR-222-3p was rejected as a normalizer due to poor detectability, failing to reach the detection threshold (Cq < 35) in 61.7% of cases (29/47). Conversely, hsa-miR-96-5p was validated as a stable reference gene. It demonstrated high detectability and expression stability (CV = 9.07%) among valid samples. Crucially, linear regression analysis showed no significant correlation between hsa-miR-96-5p levels and either the PMI (p = 0.69) and the pre-freezing time (p = 0.70). This study demonstrates that miR-222-3p is unsuitable for forensic casework in VH due to instability. We identified and validated hsa-miR-96-5p as a robust endogenous reference gene. Its adoption is recommended to standardize future molecular thanatochronology studies and improve the accuracy of PMI estimation models.

In recent years, the consumption of energy drinks by young adults and athletes has risen significantly, but concerns have been raised about the potential health risks associated with excessive consumption. These concerns include cardiovascular problems, nervous system disorders, and the potential for addiction. This review aims to examine the reported effects of acute or chronic abuse of energy drinks on human health. The analysis shows a significant prevalence of adverse effects, particularly on the cardiovascular and neurovegetative systems. In particular, the analysis identified nine cases of cardiac arrest, three of which were fatal. The aetiology of these adverse effects is attributed to the inherent neurostimulant properties of these beverages, of which caffeine is the predominant component. A comparison of documented effects in humans with experimental studies in animal models showed an overlap in results. This review highlights the need for greater rigour in the assessment of sudden cardiac death, particularly in young people, as legal substances such as energy drinks may be involved. We propose stricter limits on the consumption of these beverages than for caffeine, based on the evidence found and the data in the literature. This review also calls for the establishment of regulations governing the consumption of these products in view of their potential impact on human health.