Cross-Species Transcriptomics Analysis Highlights Conserved Molecular Responses to Per- and Polyfluoroalkyl Substances

In recent decades, per- and polyfluoroalkyl substances (PFASs) have garnered widespread public attention due to their persistence in the environment and detrimental effects on the health of living organisms, spurring the generation of several transcriptome-centered investigations to understand the biological basis of their mechanism. In this study, we collected 2144 publicly available samples from seven distinct animal species to examine the molecular responses to PFAS exposure and to determine if there are conserved responses. Our comparative transcriptional analysis revealed that exposure to PFAS is conserved across different tissues, molecules and species. We identified and reported several genes exhibiting consistent and evolutionarily conserved transcriptional response to PFASs, such as ESR1, HADHA and ID1, as well as several pathways including lipid metabolism, immune response and hormone pathways. This study provides the first evidence that distinct PFAS molecules induce comparable transcriptional changes and affect the same metabolic processes across inter-species borders. Our findings have significant implications for understanding the impact of PFAS exposure on living organisms and the environment. We believe that this study offers a novel perspective on the molecular responses to PFAS exposure and provides a foundation for future research into developing strategies for mitigating the detrimental effects of these substances in the ecosystem

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

Applications of capillary electrophoresis and high performance liquid chromatography in forensic analysis

Non disponibileForensic science include all the applications of science which can be used to support at any level the enforcement of the law. Moreover, in many countries the term forensic science also includes the scientific knowledge on the basis of which new laws and rules are drafted. Forensic analysis is a rapidly expanding field of analytical research as witnessed by the numerous publications and specific scientific reviews present in the Pubmed database, which at present includes as many as 12341 titles. It is worth noting however, that the majority of works concerning forensic analysis are related to chromatographic techniques (GC and HPLC) which can provide fast and easily comparable results, due to widespread standardized procedures. So far, little space has been given to electrophoresis. However, the low consumption of sample and reagents, the ability to separate both macromolecules and ions (both inorganic and organic) and the ruggedness of the instrumentation, have contributed to make the capillary electrophoresis (CE), known also as high-performance capillary electrophoresis (HPCE), a widely appreciated analytical technique with an extremely wild field of applications. In recent years, great attention has been paid to CE by leading forensic science laboratories at the DEA (Drug Enforcement Administration) and the FBI (Federal Bureau of Investigation). Already in 1996 CE has been discussed as an example of application in US courts of new federal rules of evidence following the Daubert standard (Kuffner et al., 1996). In this perspective, to provide further confirmation of the reliability and applicability of this technique in forensic laboratories, this work is started and aimed in its first part. The second part, instead, was aimed at confirming HPLC as a valuable tool for screening and identification and for precise and sensitive quantification of analytes of forensic interest. This subject does not need wide introduction, as probably does CE, since chromatographic techniques nowadays are commonly used for screening, identification and quantification of drugs, poisons and their metabolites, in every forensic laboratory. The areas of analytical toxicology in which chromatographic techniques are mostly used include analysis of amphetamines, cocaine, hallucinogens, opioids, anesthetics, hypnotics, benzodiazepines, antidepressants, neuroleptics, antihistamines, sulfonylurea-type antidiabetics, beta-blockers, and other cardiac drugs. So, HPLC, after long time of reluctantly, is today widely accepted in forensic analysis laboratories, whereas CE is still seen with suspicion. The aim of the present work is to show how both these techniques may be useful in the hands of forensic scientists, and particularly forensic toxicologists

Hair analysis for illicit drugs by using capillary zone electrophoresis-electrospray ionization-ion trap mass spectrometry

In forensic toxicology, hair analysis has become a well established analytical strategy to investigate retrospectively drug abuse histories. In this field, gas chromatography–mass spectrometry and high-performance liquid chromatography–mass spectrometry are currently used, often after preliminaryscreening with immunoassays. However, on the basis of previous applications to pharmaceutical analysis, capillary zone electrophoresis coupled to ion trap mass spectrometry looks also highly promising. The purpose of the present work was the development of a simple and rapid CZE–MS method for sensitive and quantitative determination of the main drugs of abuse and their metabolites (namely, 6-monoacetylmorphine, morphine, amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethampthetamine (MDMA), benzoylecgonine, ephedrine and cocaine) in human hair. Hair samples (100 mg) were washed, cut and incubated overnight in 0.1MHCl at 45 ◦C, then neutralized with NaOH and extracted by a liquid–liquid extraction method. CZE separations were carried out in a 100 cm×75m (I.D.) uncoated fused silica capillary. The separation buffer was composed of 25mM ammonium formate, pH 9.5; the separation voltage was 15 kV. Electrokineticinjections were performed at 7 kV for 30 s under field amplified sample stacking conditions. ESI-ion trap MS detection was performed in the ESI positive ionization mode using the following conditions: capillary voltage 4 kV, nebulizer gas (nitrogen) pressure 3 psi, source temperature 150 ◦C and drying gas (nitrogen) flow rate 8 l/min. A sheath liquid, composed of isopropanol–water (50:50, v/v) with 0.5% formic acid, was delivered at a flow rate of 4 ul/min. The ion trap MS operated in a selected ion monitoring mode (SIM) of positive molecular ions for each drug/metabolite.Collision induced fragmentation was also possible. Nalorphine was used as internal standard. Under the described conditions, the separation of all compounds, except amphetamine/methamphetamine, MDA/MDMA and morphine/6-MAM was achieved in 20 min, with limits of detection lower than the most severe cut-offs adopted in hair analysis (i.e. 0.1 ng/mg). Linearity was assessed within drug concentration ranges from 0.025 to 5 ng of each analyte/mg of hair. Analytical precision was fairly acceptable with RSD’s ≤3.06% for migration times and ≤22.47% for areas in realsamples, in both intra-day and day-to-day experiments. On these grounds, the described method can be proposed for rapid, selective and accurate toxicological hair analysis for both clinical and forensic purposes

Broad-spectrum toxicological analysis of hair based on capillary zone electrophoresis-time-of-flight mass spectrometry

The coupling of capillary electrophoresis\u2013electrospray ionization and time-of-flight mass spectrometry, combining efficiency and speed of separation with high mass accuracy and fast scanning capability, was for the first time applied to the determination of drugs of abuse (amphetamine, methamphetamine,MDA, MDMA, ephedrine, cocaine, morphine, codeine) and their metabolites in hair (6-MAM, benzoylecgonine). Experimental conditions were as follows. Separation: voltage 15 kV, uncoated fused-silica capillary (75 um ID, 100 cm total length), running electrolyte 25mM ammonium formate, pH 9.5, field-amplified sample stacking injection. Forensic drugs could be identified by exact mass determination (mass accuracy typically 64 5 ppm) and by match of the isotopic pattern. The method was fully validated, showing limit of detections (LODs) suitable for the determination of all the compounds below the cut-off usually adopted for hair analysis (0.1 ng/mg). Analytical precision in real matrices (tested at 0.1 and 1.0 ng/mg) was typically characterized by CV\u2019s 64 24% in both intra-day and day-to-day experiments. Quantitative determination was also tested by using a single internal standard (folcodine). Results, although with a moderate accuracy, conceivably depending on the lack of deuterated internal standards, proved useful for diagnostic use of the results from hair analysis. A single liquid\u2013liquid extraction procedure was applied for all analytes, allowing the detection of a broad spectrum of basic drugs and their major metabolites