Endurance-dependent urinary extracellular vesicle signature: shape, metabolic miRNAs, and purine content distinguish triathletes from inactive people.

Extracellular vesicles (EVs) enriched with bioactive molecules have gained considerable attention in nanotechnology because they are critical to intercellular communication while maintaining low immunological impact. Among biological matrices, urine has emerged as a noninvasive source of extracellular-contained liquid biopsy, currently of interest as a readout for physiological adaptations. Therefore, we aimed to evaluate chronic adaptations of endurance sport practice in terms of urinary EV parameters and evaluated by food consumption assessment. Two balanced groups of 13 inactive controls vs. triathlon athletes were enrolled; their urinary EVs were obtained by differential ultracentrifugation and analyzed by dynamic light scattering and transmission electron and atomic force microscopy. The cargo was analyzed by means of purine and miRNA content through HPLC-UV and qRT-PCR. Specific urinary EV signatures differentiated inactive versus endurance-trained in terms of peculiar shape. Particularly, a spheroid shape, smaller size, and lower roughness characterize EVs from triathletes. Metabolic and regulatory miRNAs often associated with skeletal muscle (i.e., miR378a-5p, miR27a-3p, miR133a, and miR206) also accounted for a differential signature. These miRNAs and guanosine in urinary EVs can be used as a readout for metabolic status along with the shape and roughness of EVs, novel informative parameters that are rarely considered. The network models allow scholars to entangle nutritional and exercise factors related to EVs' miRNA and purine content to depict metabolic signatures. All in all, multiplex biophysical and molecular analyses of urinary EVs may serve as promising prospects for research in exercise physiology.

When the Helmet Is Not Enough: Forensic Multidisciplinary Reconstruction of a Deadly Motorcycle Accident.

We report the case of a 54-year-old man who died in a motorcycle accident due to loss of control of the vehicle on a viaduct. No other vehicles were apparently involved, except for a car hit by the motorcycle after it fell. A post-mortem CT scan (computed tomography scan) was performed showing complex head trauma with a subarachnoid hemorrhage and multiple skull and facial bone fractures. A forensic cinematic reconstruction performed by an engineer was needed to exclude other incident causes other than the loss of control. The multidisciplinary approach that included autopsy findings, a cinematic reconstruction, a helmet test and an examination played a key role in clarifying the dynamics of the accident, allowing us to explain how the death occurred despite the motorcyclist's helmet use. The cause of death was identified as a penetrating head trauma with cerebral material exposure, produced by the impact of the head against a fixed bolt in the guardrail base. Despite the use of the helmet, the impact force was enough to render the protection ineffective and allowed the bolt to penetrate through the helmet and the skull.

New Perspectives for Postmortem Human Satellite Cells of Different Embryological Origin.

Human postmortem skeletal muscles are a unique source of satellite cells for skeletal muscle regenerative studies. Presomite and somite satellite cells obtained by postmortem muscles have been established as populations of human skeletal muscle precursor cells able to proliferate and differentiate in vitro. It is extremely interesting to have access to a large amount of postmortem human skeletal muscle precursor cells, especially from craniofacial as well as limb skeletal muscles in order to evaluate their potential application not only for the fundamental understanding of muscle physiology and diseases but also for drug testing in a challenging 3D-shaping muscles like skeletal muscle microphysiological systems.

Long persistence of severe acute respiratory syndrome coronavirus 2 swab positivity in a drowned corpse: a case report.

BACKGROUND: Since the beginning of the worldwide spread of severe acute respiratory syndrome coronavirus 2 to date, important knowledge has been obtained about the virus behavior in living subjects and on inanimate surfaces; however, there is still a lack of data on virus persistency on dead bodies and the risk of contagion from cadavers. CASE PRESENTATION: The present case shows the persistency of the severe acute respiratory syndrome coronavirus 2 viral genome in nasopharyngeal swabs performed on a drowned Caucasian man, aged 41 years old, who was completely asymptomatic when he was alive, up to 41 days after death. Specific real-time reverse transcriptase-polymerase chain reaction (TaqMan 2019-nCoV Assay Kit v2; Thermo Fisher Scientific, Italy and Realquality RQ-SARS-CoV-2, AB Analytical) was used to evaluate the swabs. CONCLUSIONS: This data reflect the importance of postmortem swabs in all autopsy cases, and not only in potential severe acute respiratory syndrome coronavirus 2-related death, and also highlight the necessity to evaluate virus positivity a long time after the moment of death, even if a low initial viral load was assessed.

Fast Quantitative LC-MS/MS Determination of Illicit Substances in Solid and Liquid Unknown Seized Samples.

Narcotic and psychotropic substances are natural, synthetic, or semisynthetic compounds that are present in both solid and liquid illicit products. The alterations effects on the central nervous system related to their use can be psycholeptic, psychoanaleptic, or psychodiseptic and are able to generate tolerance, addiction, or dependence phenomena, creating social and public order problems. In this scenario, the analytical evaluations that aim to determine these analytes in seized nonbiological samples, and which assume the character of judicial evidence, must meet high analytical requirements of reliability, transparency, and procedures uniformity at a national level. For the first time in the literature, the herein validated method is able to provide the simultaneous quantitative determination of 37 of the most common narcotic substances as well as the most commonly used excipients/adulterants found in seized illicit material. Additionally, the validated method can process both solid and liquid samples maintaining the precision and trueness levels (intraday and interday) in accordance with the U.S. Food and Drug Administration and European Medicines Agency international guidelines (<14.31 and <13.41%, respectively). Furthermore, it provides a simple and fast procedure for sample preparation using the dilute and shoot approach, exploiting the sensitivity and selectivity of the LC-MS/MS instrument configuration used and the signal acquisition in multiple reaction monitoring (MRM) mode (both positive and negative polarization modes).

Forensic Biochemical Markers to Evaluate the Agonal Period: A Literature Review.

Currently, forensic research is multidisciplinary with new methods and parameters useful to define the cause and time of death as well as survival/agony times. The identification of biochemical markers able to estimate agonal period has been studied by many forensic researchers. It is known that the estimation of agonal time in different types of death is not always easy, hence our interest in literature's data. The studies analyzed in this review confirm the important role of thanatobiochemistry for the estimation of survival times. Regardless of the death cause, the survival/agony time between the primary event and death influences markers concentrations in biological samples (e.g., blood, urine, cerebrospinal fluid). Different biomarkers can be used for qualitative evaluations in deaths with short and long agony (e.g., C-reactive protein, ferritin, GFAP, etc.). Instead, the quantitative interpretation showed limits due to the lack of reference cut-offs. Thanatobiochemistry is a useful tool to confirm what emerged from autopsies findings (macroscopic and histological analysis), but further studies are desirable to confirm the evidence emerging from our review of the literature.

Fabric-Phase Sorptive Membrane Array As a Noninvasive In Vivo Sampling Device For Human Exposure To Different Compounds.

This study introduces an innovative device for the noninvasive sampling and chromatographic analysis of different compounds present in exhaled breath aerosol (EBA). The new sampling device, especially in light of the recent COVID-19 pandemic that forced many countries to impose mandatory facemasks, allows an easy monitoring of the subject's exposure to different compounds they may come in contact with, actively or passively. The project combines the advantages of a fabric-phase sorptive membrane (FPSM) as an in vivo sampling device with a validated LC-MS/MS screening procedure able to monitor more than 739 chemicals with an overall analysis time of 18 min. The project involves the noninvasive in vivo sampling of the EBA using an FPSM array inserted inside an FFP2 mask. The study involved 15 healthy volunteers, and no restrictions were imposed during or prior to the sampling process regarding the consumption of drinks, food, or drugs. The FPSM array-LC-MS/MS approach allowed us to effectively exploit the advantages of the two complementary procedures (the convenient sampling by an FPSM array and the rapid analysis by LC-MS/MS), obtaining a powerful and green tool to carry out rapid screening analyses for human exposure to different compounds. The flexible fabric substrate, the sponge-like porous architecture of the high-efficiency sol-gel sorbent coating, the availability of a large cache of sorbent coatings, including polar, nonpolar, mixed mode, and zwitterionic phases, the easy installation into the facemask, and the possibility of sampling without interrupting regular activities provide FPSMs unparalleled advantages over other sampling techniques, and their applications are expected to expand to many other clinical or toxicological studies.

A fatal case of post endobronchial ultrasound transbronchial needle aspiration endobronchial haemorrhage: a case report and review of the literature.

Notwithstanding life-threatening haemorrhagic complications, endobronchial ultrasound-guided transbronchial needle aspiration represents a cornerstone in the evaluation of NSCLC patients. Due to its low invasiveness and satisfactory tolerability even in high-risk patients, it is usually scheduled in one-day surgery hospital stay. Moreover, EBUS-TBNA offers a viable alternative to other conventional endoscopic procedures such as mediastinoscopy with the addiction to gain access also to hilar nodes. We report an unexpected and rare event of post-bronchoscopy fatal endobronchial haemorrhage in a 67-year-old female patient with a right S1 solitary pulmonary nodule and concomitant multiple bilateral lymphadenopaties. According to clinical staging the patient was unfit for upfront surgery and endoscopic procedure for tissue diagnosis was scheduled. The immediate postoperative period was uneventful as no intraoperative injuries were reported. Twenty-four hours later, the patient claimed respiratory distress and, after admission to the Emergency Department, a massive and uncontrolled bleeding coming from the lower respiratory tract was diagnosed leading to a death. Although one-day surgery EBUS-TBNA is the standard of care in patients undergoing endoscopic exploration, current guidelines seem to be murky about proper case stratification in order to plan an exhaustive observation time especially in high-risk patients.

A particular case of accidental asphyxiation.

The case reported involved a 60-year-old man with psychiatric illness who was found dead at his home. He was almost naked and showed signs of death by violent asphyxiation, which led to the suspicion of homicide. Autopsy findings showed foreign material in the trachea and larynx. However, the partially digested content in his stomach, the study of the gastric content and the microscopic analysis of the airways made it possible to define this as accidental death due to inhalation of material from a diaper. Toxicological analysis detected the presence of phenytoin, valproic acid, and phenobarbital in his blood, with values within the therapeutic ranges. This is certainly a peculiar case, as particularly evident after careful review of the literature on deaths by asphyxiation in psychiatric patients. Furthermore, this study highlights the importance of a full autopsy and microscopic evaluation of any foreign bodies.

Sudden death due to catecholaminergic polymorphic ventricular tachycardia following negative stress-test outcome: genetics and clinical implications.

This paper discusses the case of a young boy who died suddenly during a football match. The victim's personal and family medical histories were negative for cardiac events. He had undergone a cardiological investigation some months before his death, enabling him to participate in competitive sports. Only post-mortem molecular analysis allowed for a clearer determination of the most plausible cause of death, which was identified as inherited arrhythmogenic heart disease, known as catecholaminergic polymorphic ventricular tachycardia. It was possible to detect a novel, previously undescribed, variant in the RYR2 gene. This case report highlights the importance of a meaningful forensic multidisciplinary investigation in such cases, and also discusses possible medical malpractice claims.

An unusual case of murder-suicide: The importance of studying knots.

An atypical murder-suicide that involved a married couple over 80 years of age was investigated at the Institute of Legal Medicine of the University of Chieti-Pescara and is presented and discussed here. The whole story allows us to confront the difficulties involved in the discovery of a dead body in water when the circumstantial evidence does not allow any unique interpretation of the facts. The atypical and misleading element appeared to be how the body of the man was tied, with the interpretation inclined towards a homicidal manner of death. It was only by combining the contributions of each of the findings that emerged during the course of the autopsy with the circumstantial evidence from the site inspection that it was possible to reach resolution of this case, which was then identified as a murder-suicide.