Molecular detection of SARS-CoV-2 from indoor air samples in environmental monitoring needs adequate temporal coverage and infectivity assessment.

The relevance of airborne exposure to SARS-CoV-2 in indoor environments is a matter of research and debate, with special importance for healthcare low-risk settings. Experimental approaches to the bioaerosol sampling are neither standardized nor optimized yet, leading in some cases to limited representativity of the temporal and spatial variability of viral presence in aerosols. Airborne viral viability moreover needs to be assessed. A study has been conducted collecting five 24-h PM10 samples in a COVID-19 geriatric ward in late June 2020, and detecting E and RdRp genes by RT-qPCR with a Ct between 36 and 39. The viral RNA detection at Ct = 36 was related to the maximal numerosity of infected patients hosted in the ward. Lacking a direct infectivity assessment for the collected samples an experimental model has been defined, by seeding twelve nasopharyngeal swab extracts from COVID-19 positive patients on Vero E6 cells; only the four extracts with a viral load above E+10 viral copies (approximately Ct<24) have been able to establish a persistent infection in vitro. Therefore, the cytopathic effect, a key feature of residual infectivity, could be considered unlikely for the environmental PM10 samples showing amplification of viral RNA at Ct = 36 or higher. A standardization of airborne SARS-CoV-2 long-term monitoring and of environmental infectivity assessment is urgently needed.

Comparison of wood smoke PM2.5 obtained from the combustion of FIR and beech pellets on inflammation and DNA damage in A549 and THP-1 human cell lines.

The aim of this study was to investigate the effect on the induction of interleukin-8 of particulate matter (PM) from fir and beech pellets burnt in domestic appliances on two human cells lines, namely the lung epithelial cell line A549 and the promyelocytic cell line THP-1. The effects of PM2.5 obtained from combustion of beech and fir pellets were compared to reference diesel exhaust particulates (DEP). In parallel, wood smoke PM-induced genotoxicity and oxidative stress were also investigated in A549 cells. Cells were treated for different times (3-72 h) with increasing concentrations of PM2.5 obtained from sequential combustions of fir and beech pellets or reference DEP. Cell viability was assessed by lactate dehydrogenase leakage, and the release of interleukin-8 or CXCL8 (IL-8) was measured to evaluate the pro-inflammatory effect. Oxidative stress was evaluated by the 5(6)-carboxy-2',7'dichlorofluorescein diacetate (DCFH-DA) assay and DNA damage by the alkaline comet assay and micronucleus frequency by flow cytometry. Both A549 and THP-1 cells responded in a dose- and time-related manner to wood smoke PM2.5 with IL-8 release, particles obtained from late combustions being the most active. THP-1 cells were more sensitive than A549 cells. On a mass base, similar effects were observed for both fir and beech PM2.5. However, the combustion of beech pellets generated approximately three times more PM2.5 than fir pellets. Regarding the mechanism of PM2.5 uptake, in both THP-1 and A549 cells, cytochalasin D prevented PM2.5-induced IL-8 mRNA expression and cytokine release, indicating a key role for actin polymerization in particles uptake and that the production of IL-8 correlated with particle phagocytosis. As signal transduction pathway involvement, in both THP-1 and A549 cells, PM2.5-induced IL-8 release could be completely blocked by the selective inhibitor SB203580, indicating a role of p38 MAPK activation. PM2.5 from both fir and beech pellets also induced modest DNA lesions dose related, measured as strand breaks, whereas no increase in the number of micronucleus was observed. Similar effects were observed with DEP, arguing against less dangerous effects of wood smoke particles than other categories of combustion-derived particles in the same size range. Overall, results suggest that combustion conditions can significantly affect the characteristics of particles and the consequent toxicity, and that different woods can generate different amounts of PM2.5.

Urban BTEX Spatiotemporal Exposure Assessment by Chemometric Expertise.

Normative regulations on benzene in fuels and urban management strategies are expected to improve air quality. The present study deals with the application of self-organizing maps (SOMs) in order to explore the spatiotemporal variations of benzene, toluene, ethylbenzene, and xylene levels in an urban atmosphere. Temperature, wind speed, and concentration values of these four volatile organic compounds were measured after passive sampling at 21 different sampling sites located in the city of Trieste (Italy) in the framework of a multi-year long-term monitoring program. SOM helps in defining pollution patterns and changes in the urban context, showing clear improvements for what concerns benzene, toluene, ethylbenzene, and xylene concentrations in air for the 2001-2008 timeframe.

Non-invasive assessment of exhaled breath pattern in patients with multiple chemical sensibility disorder.

Multiple chemical sensitivity (MCS) is a complex disorder initiated by chemical exposure, particularly through the airways. MCS patients report sensitivity or intolerance to low levels of a wide spectrum of chemicals. Symptoms could include asthma-like signs, rhinitis, fatigue, cognitive dysfunction, psycho-physiological alteration, and other specific tissue reactions resembling hypoxic and oxidative stress effects. To recognize physiological signs that would allow the diagnosis of MCS in a non-invasive way we investigated the potential application of a new sensor system. In healthy volunteers, we measured exhaled breath content in the control condition and under exposure to olfactory stressors that mimic hypoxic or pollutant stressors playing a potential role in the generation of the MCS disorder. The recording system used is based on metal oxide semiconductor (MOS) sensor having a sensing range of 450-2,000 ppm CO(2) equivalents, which is able to detect a broad range of compounds playing a potential role in the generation of the MCS disorder, while correlating directly with the CO(2) levels. The results indicate that the recording system employed was suitable for the analysis of exhaled breath content in humans. Interestingly, the system was able to detect and discriminate between the exhaled breath content taken from the control condition and those from conditions under stress that mimicked exposures to pollutant or hypoxia. The results suggest that chronic hypoxia could be involved in the MCS disorder.

[Particulate matter and atmospheric pollutants exposure for outdoor workers in the Muggia area (Trieste, Italy)]. / Indagine sull'esposizione a polveri ed inquinanti atmosferici di lavoratori outdoor nell'area di Muggia (Trieste).

UNLABELLED: Urban air pollution originates from traffic, heating and industrial activities and it exposes several categories of workers to noxious chemicals; in recent years various surveys have been carried on taxi drivers, bus drivers, policemen, newspaper kiosks, fuel station workers and general population, for assessing exposure to main contaminants, benzene and PAHs in particular. AIM: defining a specific sampling strategy for collecting an experimental data base for the assessment of potential population exposure to atmospheric particulate matter in the Muggia area (Trieste, North-East Italy). MATERIALS AND METHODS: A sample of 14 urban police officers working mainly on the roads and potentially subject to occupational exposure has been analyzed, as well as a sample of the municipal administrative personnel (13 people) working in indoor environments during the same sampling periods. Exposure has been monitored by personal samplers and high volume environmental samples; urinary levels of 1 OH-pyrene and creatinine were collected for each participant at the end of each sampling period. RESULTS: PAHs analyses provided diagnostic ratios for identification of the sources of pollution. In general experimental BaP values stay below the environmental target value, with satisfactory agreement between environmental and personal monitoring. Levels of urinary 1 OH-pyrene for administrative personnel was detected always below the background value (0.089 micromoles/mole creatinine), while they are above the background and below the attention value for most of urban police officers. CONCLUSIONS: Municipal police officers participating to the present study, while belonging to one of the categories of people most exposed to outdoor PM, do not show any worrying exposure to the considered chemicals associated to their occupational activity, in comparison to other factors, possibly associated to personal lifestyles.