Congenital heart diseases (CHDs) and forensic investigations: Searching for the cause of death.

Congenital Heart Diseases (CHDs) are a group of structural abnormalities or defects of the heart that are present at birth. CHDs could be connected to sudden death (SD), defined by the WHO (World Health Organization) as "death occurring within 24 h after the onset of the symptoms" in an apparently "healthy" subject. These conditions can range from relatively mild defects to severe, life-threatening anomalies. The prevalence of CHDs varies across populations, but they affect millions of individuals worldwide. This article aims to discuss the post-mortem investigation of death related to CHDs, exploring the forensic approach, current methodologies, challenges, and potential advancements in this challenging field. A further goal of this article is to provide a guide for understanding these complex diseases, highlighting the pivotal role of autopsy, histopathology, and genetic investigations in defining the cause of death, and providing evidence about the translational use of autopsy reports. Forensic investigations play a crucial role in understanding the complexities of CHDs and determining the cause of death accurately. Through collaboration between medical professionals and forensic experts, meticulous examinations, and analysis of evidence, valuable insights can be gained. These insights not only provide closure to the families affected but also contribute to the prevention of future tragedies.

Nicotine dosimetry and stability in cambridge filter PADs (CFPs) following different smoking regime protocols and storage conditions.

Despite the growing numbers of studies on cigarettes and electronic nicotine delivery products (ENDs), no standard assessment of nicotine stability in various matrix post exposure is currently available. The aim of the present study was to evaluate the optimal standard condition to store Cambridge Filter Pads (CFPs) before chemical analysis in order to guarantee the titer of nicotine.We further performed data normalization according to different smoking or vaping runs. Smoke and vapor generated respectively by a reference tobacco cigarette (1R6F) and ENDs under different exposure regimes (ISO, HCI and CRM81) were collected on CFPs as total particulate matter (TPM) and subsequently analyzed for nicotine content. For each exposure, some CFPs were analyzed at time zero, whereas the others were stored under different conditions for nicotine assessment after 30 days. Principal Component Analysis (PCA) showed the best correlation between nicotine on CFPs and TPM for normalization. This study suggests that different exposure regimes and products can affect the preservation of nicotine titer on CFPs while samples storage at -80 °C may prevent the loss of nicotine. Finally, normalization of nicotine with TPM is strongly recommended for regulatory purpose.

The role of air pollution (PM and NO2) in COVID-19 spread and lethality: A systematic review.

A new coronavirus (SARS-CoV-2) has determined a pneumonia outbreak in China (Wuhan, Hubei Province) in December 2019, called COVID-19 disease. In addition to the person-to person transmission dynamic of the novel respiratory virus, it has been recently studied the role of environmental factors in accelerate SARS-CoV-2 spread and its lethality. The time being, air pollution has been identified as the largest environmental cause of disease and premature death in the world. It affects body's immunity, making people more vulnerable to pathogens. The hypothesis that air pollution, resulting from a combination of factors such as meteorological data, level of industrialization as well as regional topography, can acts both as a carrier of the infection and as a worsening factor of the health impact of COVID-19 disease, has been raised recently. With this review, we want to provide an update state of art relating the role of air pollution, in particular PM2.5, PM10 and NO2, in COVID-19 spread and lethality. The Authors, who first investigated this association, often used different research methods or not all include confounding factors whenever possible. In addition, to date incidence data are underestimated in all countries and to a lesser extent also mortality data. For this reason, the cases included in the reviewed studies cannot be considered conclusive. Although it determines important limitations for direct comparison of results, and more studies are needed to strengthen scientific evidences and support firm conclusions, major findings are consistent, highlighting the important contribution of PM2.5 and NO2 as triggering of the COVID-19 spread and lethality, and with a less extent also PM10, although the potential effect of airborne virus exposure it has not been still demonstrated.

Micro- and nano-plastics in edible fruit and vegetables. The first diet risks assessment for the general population.

Microplastics (MPs) represent a current public health concern since toxicity has not yet fully investigated. They were found in several foods, but to the best of our knowledge, at this time no data was reported for the edible vegetables and fruits. We focused on diet exposure aiming to evaluate the number and the size (<10 µm) of MPs in the most commonly consumed vegetables and fruits, in relation to their recommended daily intake too. MPs extraction and analysis were carried out using an innovative Italian methodology and SEM-EDX, respectively. Finally, we calculated the Estimated Daily Intakes (EDIs) for adults and children for each type of vegetal and fruit. The higher median (IQR) level of MPs in fruit and vegetable samples was 223,000 (52,600-307,750) and 97,800 (72,175-130,500), respectively. In particular, apples were the most contaminated fruit samples, while carrot was the most contaminated vegetable. Conversely, the lower median (IQR) level was observed in lettuce samples 52,050 (26,375-75,425). Both vegetable and fruit samples MPs levels were characterized by wide variability. The smallest size of MPs was found in the carrot samples (1.51 µm), while the biggest ones were found in the lettuce (2.52 µm). Both vegetable and fruit samples had size of the MPs characterized by low variability. We found the highest median level of MPs in samples purchased from the "fruiter 3" (124,900 p/g) and the lowest in those purchased in "supermarket" (87,600 p/g). The median size of the MPs had overlapping dimensions in all the purchase sites, with the exception of the samples purchased at the "shop at km zero 2″ which had slightly smaller size (1.81 µm). The highest adults' (4.62 E+05) and children's (1.41 E+06) EDIs are due the ingestion of apples, instead the lowest are due to the ingestion of carrots (adults: 2.96 E+04; children: 1.15 E+05). We hypothesized that the mechanism of uptake and translocation of MPs can be the same described and reported for carbon-nanomaterials. This may be a possible translocation route of MPs by environment to vegetables permitting, so, the translocation or uptake inside of their biological systems. Based on the results obtained it is urgent important to perform toxicological and epidemiological studies to investigate for the possible effects of MPs on human health.

Phytoremediation potential of Arundo donax (Giant Reed) in contaminated soil by heavy metals.

Soil pollution from heavy metals poses a serious risk for environment and public health. Phytoremediation is an eco-friendly and cheaper alternative compared to chemical-physical techniques. We carried out in vitro tests where three microorganisms Trichoderma harzianum, Saccharomyces cerevisiae and Wickerhamomyces anomalus were exposed to eight different heavy metals (one metal at a time) in order to evaluate resistance, growth and bioaccumulation capability for each metal (Ni, Cd, Cu, V, Zn, As, Pb, Hg). Taking into account the natural characteristics of T. harzianum, (resistance to environmental stress, resistance to pathogenic fungi, ability to establish symbiotic relationships with superior green plants) and the good bioaccumulation capacity for V, As, Cd, Hg, Pb shown after in vitro tests, it was chosen as a microorganism to be used in greenhouse tests. Controlled exposure tests were performed in greenhouse, where Arundo donax and mycorrhized Arundo donax with T. harzianum were exposed for 7 months at two different doses (L1 and L2) of a heavy metal mix, so as to assess whether the symbiotic association could improve the bioaccumulation capability of the superior green plant A. donax. Heavy metals were determined with ICP-MS. The average bioaccumulation percentage values of A. donax for L1 and L2 were, respectively: Ni (31%, 26%); Cd (35%, 50%); Cu (30%, 35%); As (19%, 27%); Pb (18%, 14%); Hg (42%, 45%); V (39%, 26%); Zn (23%, 9%). The average bioaccumulation percentage values of mycorrhized A. donax with T. harzianum for L1 and L2 were, respectively: Ni (27%, 38%); Cd (44%, 42%); Cu (36%, 29%); As (17%, 23%); Pb (37%, 54%); Hg (44%, 60%); V (16%, 20%); Zn (14%, 7%). A. donax showed the highest BAF (bioaccumulation factor) for Cd (0.50), Cu (0.35), As (0.27) and Hg (0.45) after exposure to L2; mycorrhized A. donax with T. harzianum showed the highest BAF for Ni (0.38), Cd (0.42), Pb (0.54) and Hg (0.60) after exposure to L2. A. donax showed the highest TF (translocation factor) values for Cd (0.28) and Hg (0.26) after exposition at L1 and L2 respectively; A. donax mycorrhized with T. harzianum showed the highest TF values for Cd (0.70), As (0.56), V (0.24), Pb (0.18) after exposition at L2, and Zn (0.30) after exposition at L1. Our study showed a good growth capability in contaminated soils and a good bioaccumulation capability of heavy metals, both for A. donax and mycorrhized A. donax with T. harzianum. Furthermore, for three metals (Ni, Pb and Hg) the bioaccumulation capability was improved by the symbiosis of T. harzianum with A. donax. So, these results proved the suitability both for A. donax and mycorrhized A. donax with T. harzianum for phytoremediation processes.

Living near waterbodies as a proxy of cyanobacteria exposure and risk of amyotrophic lateral sclerosis: a population based case-control study.

BACKGROUND: Epidemiological studies highlighted the possibility that exposure to cyanotoxins leads to the development of the neurodegenerative disease amyotrophic lateral sclerosis (ALS). METHODS: We devised a population-based case-control study in two Italian populations. We used residential proximity of the residence to water bodies as a measure of possible exposure to cyanotoxins. RESULTS: Based on 703 newly-diagnosed ALS cases and 2737 controls, we calculated an ALS odds ratio (OR) of 1.41 (95% CI: 0.72-2.74) for current residence in the vicinity of water bodies, and a slightly lower estimate for historical residence (OR: 1.31; 95% CI: 0.57-2.99). Subjects <65 years and people living in the Northern Italy province of Modena had higher ORs, especially when historical residence was considered. CONCLUSIONS: Overall, despite some risk of bias due to exposure misclassification and unmeasured confounding, our results appear to support the hypothesis that cyanotoxin exposure may increase ALS risk.

[Algal biotoxins in Dialysis Water: a risk not managed]. / Biotossine algali in acque di dialisi: un rischio non gestito.

A literature review was performed to retrieve updated information on the quality of dialysis water, with a focus on the emerging problem of the presence of algal toxins (microcystins) produced by cyanobacteria. Current legislation was examined as well as studies conducted to date in different geographic areas. In this article, the authors present review results along with recommendations to operators and managers of dialysis units, for preventing possible risks for patients.

Development and validation of a LC-MS/MS method for analysis of vortioxetine in postmortem specimens. First data from an authentic case.

Vortioxetine is an antidepressant recently licensed in USA and EU for the treatment of major depressive disorder. Neither fatal case due to overdose nor data about postmortem concentrations on blood or other specimens have been reported. The aims of this study were the development and validation of a method for vortioxetine analysis by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) in postmortem samples and its application in an authentic case. The method was validated and applied on blood, vitreous humor, bile, brain, liver, kidney, and gastric content. After protein precipitation, the supernatant was directly injected into LC-MS/MS. Analysis was carried out by Multiple Reaction Monitoring (MRM) mode. The authentic case concerned a 38 years-old woman, affected by depression, who was found hanged at home. The method determined an acceptable sensitivity, selectivity, linearity, precision, and accuracy for all matrices. No interference was shown for all matrices. The matrices do not significantly reduce the peak intensity of vortioxetine. No carryover was shown. Toxicological analysis of the authentic case showed vortioxetine in blood (234 ng/ml), vitreous humor (10.5 ng/ml), brain (490 ng/g), lung (479 ng/g), liver (3751 ng/g), kidney (798 ng/g), bile (2267 ng/ml) and gastric content (253 ng/ml). Our case suggests that even at blood concentrations of vortioxetine equal to 234 ng/ml, the subject was able to stage and carry out the hanging. Vortioxetine concentrations found in the other cadaveric samples (biological fluids, organs, and gastric content) may be helpful to evaluate further similar comparable cases.

Advances in Technologies in Crime Scene Investigation.

Crime scene investigation (CSI) is the complex act of reconstructing the dynamics that led to a crime and the circumstances of its perpetration. Crystallizing the CSI is a difficult task for the forensic pathologist; however, it is often requested by the public prosecutor and many judicial cases remain unsolved precisely for this reason. Recent years have seen an improvement in the ability of 3D scanning technology to obtain dense surface scans of large-scale spaces, for surveying, engineering, archaeology, and medical purposes such as forensics. The applications of this new technology are growing every day: forensic measurement of wounds in clinical reports, for example, reconstruction of traffic accidents, bullet trajectory studies in gunshot wounds, and 3D bloodstain pattern analysis. A retrospective analysis was conducted across all crime scene investigations performed by the forensic staff of the Department of Forensic Pathology of the University of Catania from January 2019 to June 2022. Inclusion criteria were the use of a laser scanner (LS), the use of a camera, a full investigative scene, and collection of circumstantial data thanks to the help of the judicial police. Cases in which the LS was not used were excluded. Out of 200 CSIs, 5 were included in the present study. In case number 1, the use of the LS made it possible to create a complete scale plan of the crime scene in a few hours, allowing a ship to be quickly returned to the judicial police officer. In case 2 (fall from a height), the LS clarified the suicidal intent of the deceased. In case number 3 it was possible to reconstruct a crime scene after many years. In case 4, the LS provided a great contribution in making a differential diagnosis between suicide and homicide. In case 5, the LS was fundamental for the COVID team in planning the study of COVID-19 pathways and areas within a hospital with the aim of reduction of nosocomial transmission. In conclusion, the use of the LS allowed the forensic staff to crystallize the investigative scene, making it a useful tool.

Systematic Review of Penetrating Cardiac Injury by a Firearm: Forensic Implications.

Penetrating injuries of the heart, named penetrating cardiac injury (PCI), may cause hemorrhagic shock as well as cardiac tamponade, leading to death if not treated immediately. This systematic review aims to highlight the main aspects of penetrating cardiac injuries after firearm wounds. The cases of 39 subjects (age 37.05 + 15.4) were selected (6 fatal cases). Specifically, 4/39 cases involved subjects under 18 y.o.; analyzing the entrance wound, in 30/39 cases it was located in the anterior chest, 4/39 in the posterior chest, 3/39 in the shoulder/axilla area, 1/39 in the neck, and 1/39 in the pelvis (gluteus). The exit wound was found in only 3/39 cases. Several factors may influence the prognosis: firstly, prompt intervention represents a crucial point, then considering the complications related to PCI, the most important are myocardial infarction, and projectile migration with embolization. The mortality rate is related to: (1) area and severity of the heart injury; (2) duration of transport and intervention; (3) contemporary lesion to other organ/s; (4) the quantity of blood lost; (5) and presence/absence of cardiac tamponade. Based on these findings, a correct approach in the management of PCI may be considered important from a forensic point of view, both as regards to medical liability and from the trial perspective.

In vitro cytoxicity profile of e-cigarette liquid samples on primary human bronchial epithelial cells.

Cigarette smoke is associated to severe chronic diseases. The most harmful components of cigarette smoke derive from the combustion process, which are significantly reduced in the electronic cigarette aerosol, thus providing a valid option in harm reduction strategies. To develop safer products, it is therefore necessary to screen electronic cigarette liquids (e-liquids) to meet high safety standards defined by government regulations. The aim of the present study was to evaluate the presence of metal- and plastic-derived contaminants in four different commercial e-liquids with high concentration of nicotine and their cytotoxic effect in normal human bronchial epithelial cells by a number of in vitro assays, in comparison with the 1R6F reference cigarette, using an air-liquid interface (ALI) exposure system. Moreover, we evaluated the effect of aerosol exposure on oxidative stress by measuring the production of reactive oxygen species and mitochondrial potential. Our results showed no contaminants in all e-liquids and a significantly reduced cytotoxic effect of e-liquid aerosol compared to cigarette smoke as well as a maintained mitochondria integrity. Moreover, no production of reactive oxygen species was detected with e-cigarette aerosol. In conclusion, these results support the reduced toxicity potential of e-cigs compared to tobacco cigarettes in an in vitro model resembling real life smoke exposure.

Comparative assessment of electronic nicotine delivery systems aerosol and cigarette smoke on endothelial cell migration: The Replica Project.

Cigarette smoking is associated with impairment of repair mechanisms necessary for vascular endothelium homeostasis. Reducing the exposure to smoke toxicants may result in the mitigation of the harmful effect on the endothelium and cardiovascular disease development. Previous investigations evaluated in vitro the effect of electronic cigarette (EC) compared with cigarette smoke demonstrating a significant reduction in human umbilical vein endothelial cells (HUVECs) migration inhibition following EC aerosol exposure. In the present study, we replicated one of these studies, evaluating the effects of cigarette smoke on endothelial cell migration compared with aerosol from EC and heated tobacco products (HTPs). We performed an in vitro scratch wound assay on endothelial cells with a multi-center approach (ring-study) to verify the robustness and reliability of the results obtained in the replicated study, also testing the effect of aerosol from two HTPs on endothelial cells. Consistently with the original study, we observed a substantial reduction of the effects of aerosol from EC and HTPs on endothelial cell migration compared with cigarette smoke. While cigarette smoke reduced endothelial wound healing ability already at low concentrations (12.5%) and in a concentration-dependent manner, EC and HTPs aerosol showed no effect on endothelial cells until 80%-100% concentrations. In conclusion, our study further confirms the importance of EC and tobacco heated products as a possible harm reduction strategy for cardiovascular diseases development in smokers.

Diagnosis of lethal cyanide poisoning. Analysis by Anion-Exchange Chromatography with Pulsed Amperometric Detection.

Cyanide is a poison widely used in cases of suicide or homicide. Although various methods to identify and quantify this substance are reported in the literature, they are mainly validated on biological fluids (e.g., blood and urine). In the present study, the Anion-Exchange Liquid Chromatography with Pulsed Amperometric Detection (IC-PAD) method was validated on blood and, for the first time, on gastric content, and organs (brain, lung, and liver). For each matrix, linearity, accuracy, precision, limit of detection (LOD), lower limit of quantification (LLOQ), matrix interferences, and carryover were assessed. The samples were extracted by steam distillation in acid environment for the following analysis by IC-PAD. Furthermore, cyanide values found in two real poisoning cases are reported. For each investigated matrix, the analytical method satisfied all acceptance criteria for validation: it showed a good precision and accuracy, selectivity, and sensitivity with no carryover and matrix interference. The extraction by steam distillation in acid environment REDUCED the interference of the matrices and ALLOWED to perform the analysis with good precision and accuracy. In case #1, analysis showed a blood cyanide concentration of 0.99 µg/ml. In case #2, cyanide concentrations were 1.3 µg/g in brain, 0.8 µg/g in lung, 1.6 µg/g in liver, and 1.2 µg/g in gastric content. The cyanide concentrations found in the two reported cases have been suitable to cause death by poisoning.

Heart rupture as an acute complication of cocaine abuse: A case report.

The purpose of this study is to show a very rare complication of acute cocaine poisoning, namely heart rupture. In the present case report, acute cocaine intoxication caused massive myocardial infarction, resulting in heart rupture and cardiac tamponade. A crime scene investigation found a dead body on the street in a drug dealing district. Examination of the body showed no external injuries. A thorough autopsy was performed showing massive cardiac tamponade with 510 ml of blood within the pericardium and full-thickness tissue lesion at the posterior wall of the left ventricle of 3.5 × 3 cm. Histological examination in hematoxylin and eosin was performed and confirmed the interruption of the posterior wall of the left ventricle with the presence of blood. In fact, although the correlation between cocaine and myocardial damage is well established, the relationship between heart rupture and acute cocaine intoxication is an extremely rare event. Moreover, since there are, to date, few reports of similar deaths, our report provides useful information regarding sudden death in a cocaine abuser. It is, therefore, of crucial importance to report this case to the scientific community.

The effect of laboratory-verified smoking on SARS-CoV-2 infection: results from the Troina sero-epidemiological survey.

Previous research yielded conflicting results on the association between cigarette smoking and risk of SARS-CoV-2 infection. Since the prevalence of smoking is high globally, the study of its impact on COVID-19 pandemic may have considerable implications for public health. This study is the first to investigate the association between the SARS-CoV-2 antibody sero-positivity and biochemically verified smoking status, to refine current estimates on this association. SARS-CoV-2-specific IgG and serum cotinine levels (a well-known marker of tobacco exposure) were assessed in a large sero-epidemiological survey conducted in the town of Troina (Sicily, Italy). A propensity score matching was carried out to reduce the effect of possible factors on SARS-CoV-2 infection risk among study participants. Of the 1785 subjects included in our study, one-third was classified as current smokers, based on serum cotinine levels. The overall proportion of subjects with positive serology for SARS-CoV-2 IgG was 5.4%. The prevalence of SARS-CoV-2 antibody positivity and previous COVID-19 diagnosis were reduced in smokers. This reduced prevalence persisted after adjusting for possible confounders (such as sex, age, previous infection, chronic conditions, and risk group) at regression analyses, and the point estimates based on the PS-matched models resulted consistent with those for the unmatched population. This study found a lower proportion of positive SARS-CoV-2 serology among current smokers, using direct laboratory measures of tobacco exposure and thus avoiding possible bias associated with self-reported smoking status. Results may also serve as a reference for future clinical research on potential pharmaceutical role of nicotine or nicotinic-cholinergic agonists against COVID-19.

Development and Validation of a Method for the Detection and Quantification of Antidepressants and Antipsychotics in Bone Samples by GC-MS.

Bone marrow is the tissue contained inside the bones and can be considered one of the potential alternative tissues in forensic toxicology. This matrix could be particularly useful in those cases where the routine sample is not available due to an advanced state of decomposition or skeletonization of the corpse. The aim of this study was to develop, validate and apply an analytical method of extraction and analysis of different antidepressants and antipsychotics, commonly used in therapy, from spiked pig ribs. Pig ribs, each of 5 g and 5 cm long, were spiked at three concentration levels (100, 200 and 500 ng/g). For each concentration, 10 pig ribs were prepared. The method involves the leaching by ethanol at different pHs of nine drugs from the inside of ribs, in particular from the bone marrow, without the fragmentation of bone tissue. Following a liquid-liquid purification and extraction, analysis was performed by gas chromatography coupled with mass spectrometry in selected ion monitoring mode. For method validation was assessed linearity, sensitivity, precision and accuracy, matrix interferences and, finally, carryover. Analytical method performance was acceptable respect to acceptance criteria for validation. No matrix interferences were detected; for this reason, it is possible to affirm that this method has a good selectivity. Moreover, the method was not affected by carryover. Considering that the study conducted on pig ribs has given encouraging results, it can be assumed that this method can be used in forensic toxicological protocols (human autopsy cases) as alternative to classic procedures.

Chemical Characterization and Quantification of Silver Nanoparticles (Ag-NPs) and Dissolved Ag in Seafood by Single Particle ICP-MS: Assessment of Dietary Exposure.

This study provides a first insight on the chemical characterization and quantification of silver nanoparticles (AgNPs) and dissolved Ag in processed canned seafood products, where food-grade edible silver (E174) is not intentionally added nor is the nanoparticle contained in the food contact material. The aim was to evaluate the bioaccumulation potential of AgNPs and to contribute to the assessment of AgNPs and ionic Ag human dietary intake from processed seafood. It is known how seafood, and in particular pelagic fish, is a precious nutritional source of unsaturated fatty acids, protein, and different micronutrients. Nevertheless, it may cause possible health problems due to the intake of toxic compounds coming from environmental pollution. Among emerging contaminants, AgNPs are widely applied in several fields such as biomedicine, pharmaceutical, food industry, health care, drug-gene delivery, environmental study, water treatments, and many others, although its primary application is in accordance with its antimicrobial property. As a consequence, AgNPs are discharged into the aquatic environment, where the colloidal stability of these NPs is altered by chemical and physical environmental parameters. Its toxicity was demonstrated in in-vitro and in-vivo studies, although some findings are controversial because toxicity depends by several factors such as size, concentration, chemical composition, surface charge, Ag+ ions released, and hydrophobicity. The new emerging technique called single-particle inductively coupled plasma mass spectrometry (spICP-MS) was applied, which allows the determination of nanoparticle number-based concentration and size distribution, as well as the dissolved element. Our findings highlighted comparable mean sizes across all species analysed, although AgNPs concentrations partly follow a trophic level-dependent trend. The low mean size detected could be of human health concern, since, smaller is the diameter higher is the toxicity. Dietary intake from a meal calculated for adults and children seems to be very low. Although seafood consumption represents only a small part of the human total diet, our findings represent a first important step to understand the AgNPs dietary exposure of the human population. Further studies are needed to characterize and quantify AgNPs in a large number of food items, both processing and not, and where AgNPs are added at the industrial level. They will provide a realistic exposure assessment, useful to understand if AgNPs toxicity levels observed in literature are close to those estimable through food consumption and implement data useful for risk assessors in developing AgNPs provisional tolerable daily intake.

Role of Cigarette Smoke on Angiotensin-Converting Enzyme-2 Protein Membrane Expression in Bronchial Epithelial Cells Using an Air-Liquid Interface Model.

Prevalence studies of current smoking, among hospitalized COVID-19 patients, demonstrated an unexpectedly low prevalence among patients with COVID-19. The aim of the present study was to evaluate the effect of smoke from cigarettes on ACE-2 in bronchial epithelial cells. Normal bronchial epithelial cells (H292) were exposed to smoke by an air-liquid-interface (ALI) system and ACE-2 membrane protein expression was evaluated after 24 h from exposure. Our transcriptomics data analysis showed a significant selective reduction of membrane ACE-2 expression (about 25%) following smoking exposure. Interestingly, we observed a positive direct correlation between ACE-2 reduction and nicotine delivery. Furthermore, by stratifying GSE52237 as a function of ACE-2 gene expression levels, we highlighted 1,012 genes related to ACE-2 in smokers and 855 in non-smokers. Furthermore, we showed that 161 genes involved in the endocytosis process were highlighted using the online pathway tool KEGG. Finally, 11 genes were in common between the ACE-2 pathway in smokers and the genes regulated during endocytosis, while 12 genes with non-smokers. Interestingly, six in non-smokers and four genes in smokers were closely involved during the viral internalization process. Our data may offer a pharmaceutical role of nicotine as potential treatment option in COVID-19.

Seroepidemiological Survey on the Impact of Smoking on SARS-CoV-2 Infection and COVID-19 Outcomes: Protocol for the Troina Study.

BACKGROUND: After the global spread of SARS-CoV-2, research has highlighted several aspects of the pandemic, focusing on clinical features and risk factors associated with infection and disease severity. However, emerging results on the role of smoking in SARS-CoV-2 infection susceptibility or COVID-19 outcomes are conflicting, and their robustness remains uncertain. OBJECTIVE: In this context, this study aims at quantifying the proportion of SARS-CoV-2 antibody seroprevalence, studying the changes in antibody levels over time, and analyzing the association between the biochemically verified smoking status and SARS-CoV-2 infection. METHODS: The research design involves a 6-month prospective cohort study with a serial sampling of the same individuals. Each participant will be surveyed about their demographics and COVID-19-related information, and blood sampling will be collected upon recruitment and at specified follow-up time points (ie, after 8 and 24 weeks). Blood samples will be screened for the presence of SARS-CoV-2-specific antibodies and serum cotinine, being the latter of the principal metabolite of nicotine, which will be used to assess participants' smoking status. RESULTS: The study is ongoing. It aims to find a higher antibody prevalence in individuals at high risk for viral exposure (ie, health care personnel) and to refine current estimates on the association between smoking status and SARS-CoV-2/COVID-19. CONCLUSIONS: The added value of this research is that the current smoking status of the population to be studied will be biochemically verified to avoid the bias associated with self-reported smoking status. As such, the results from this survey may provide an actionable metric to study the role of smoking in SARS-CoV-2 infection and COVID-19 outcomes, and therefore to implement the most appropriate public health measures to control the pandemic. Results may also serve as a reference for future clinical research, and the methodology could be exploited in public health sectors and policies. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/32285.

Electronic nicotine delivery systems exhibit reduced bronchial epithelial cells toxicity compared to cigarette: the Replica Project.

Electronic nicotine delivery systems (ENDS) may reduce health risks associated with chronic exposure to smoke and their potential benefits have been the matter of intense scientific debate. We aimed to replicate three published studies on cytotoxic and inflammatory effects of cigarette smoke and ENDS aerosol in an independent multi-center ring study. We aimed to establish the reliability of results and the robustness of conclusions by replicating the authors' experimental protocols and further validating them with different techniques. Human bronchial epithelial cells (NCI-H292) were exposed to cigarette whole smoke and vapor phase and to aerosol from ENDS. We also assessed the inflammatory cytokines interleukin-6 and interleukin-8 and the remodeling mediator matrix metalloproteinase-1. We replicated cell viability results and confirmed that almost 80% of cytotoxic effects are due to volatile compounds in the vapor phase of smoke. Our findings substantiated the reduced cytotoxic effects of ENDS aerosol. However, our data on inflammatory and remodeling activity triggered by smoke differed significantly from those in the original reports. Taken together, independent data from multiple laboratories clearly demonstrated the reduced toxicity of ENDS products compared to cigarettes.