Rotenone-induced oxidative stress in THP-1 cells: biphasic effects of baicalin.

BACKGROUND: Several results demonstrated that microglia and peripheral monocytes/macrophages infiltrating the central nervous system (CNS) are involved in cell response against toxic compounds. It has been shown that rotenone induces neurodegeneration in various in vitro experimental models. Baicalin, a natural compound, is able to attenuate cell damage through anti-oxidant, anti-microbial, anti-inflammatory, and immunomodulatory action. Using THP-1 monocytes, we investigated rotenone effects on mitochondrial dysfunction and apoptosis, as well as baicalin ability to counteract rotenone toxicity. METHODS AND RESULTS: THP-1 cells were exposed to rotenone (250 nM), in the presence/absence of baicalin (10-500 µM) for 2-24 h. Reactive Oxygen Species production (ROS), mitochondrial activity and transmembrane potential (Δψm), DNA damage, and caspase-3 activity were assessed. Moreover, gene expression of mitochondrial transcription factor a (mtTFA), interleukin-1ß (IL-1ß), B-cell lymphoma 2 (Bcl2) and BCL2-associated X protein (Bax), together with apoptotic morphological changes, were evaluated. After 2 h of rotenone incubation, increased ROS production and altered Δψm were observed, hours later resulting in DNA oxidative damage and apoptosis. Baicalin treatment at 50 µM counteracted rotenone toxicity by modulating the expression levels of some proteins involved in mitochondrial biogenesis and apoptosis. Interestingly, at higher baicalin concentrations, rotenone-induced alterations persisted. CONCLUSIONS: These results give evidence that exposure to rotenone may promote the activation of THP-1 monocytes contributing to enhanced neurodegeneration. In this context, baicalin at low concentration exerts beneficial effects on mitochondrial function, and thus may prevent the onset of neurotoxic processes.

The Italian Mandatory Notification System: An Important Public Health Tool For Continuous Monitoring Of Infectious Diseases.

Infectious diseases still register significant morbidity and mortality worldwide. Surveillance through a mandatory notification system allows the continuous analysis of the situation even at a local level and its importance has been highlighted by the recent COVID-19 pandemic. This paper aimed to outline the importance of the mandatory notification system as a Public Health tool in the continuous monitoring of infectious diseases. To this aim, we carried out a cross-sectional study examining the notifications reported in the Italian territory of Messina, Sicily, in the period 2001-2020. The institutional websites were examined and the notification data were used to obtain the incidences. Overall, a significant reduction of the incidence notification trend was observed. Chickenpox was by far the most notified infectious disease, followed by scabies, pediculosis, and brucellosis. Outbreaks of brucellosis, measles and hepatitis A occurred. All the diseases decreased over time, except syphilis, for which a significant increase was observed. Surveillance of infectious diseases through a mandatory notification system remains a bulwark of public health despite underreporting. Our study reflects the situation of a typical high-income area, although some unexpected criticisms are highlighted. Continuous information about correct behaviors through education campaigns are crucial in order to improve the situation. Keywords: mandatory notifications, infectious diseases, surveillance, public health Corresponding author: Alessio Facciolà, Department of Biomedical and Dental Sciences and Morphofunctional Imaging, University of Messina, Italy. Email: afacciola@unime.it.

Prevalence of HBsAg and HCV antibodies in a cohort of pregnant women: the continuous survey as public health tool against pregnant and neonatal complications.

Pregnancy is a very delicate condition and different external factors can alter fetal development. Microorganisms are surely the principal ones. Several studies have shown that HBV and HCV infections could have an impact on pregnancy outcome. We performed a survey of blood samples from pregnant women in order to evaluate the presence of HBsAg and HCV antibodies. 6,896 women were tested from 2016 to 2019. 0.2% of the women in the Italian group and 2.1% among the foreigners tested positive for HBsAg, while 0.2% among the Italians and 0.7% among the foreigners were positive for HCV antibodies. Moreover, an increasing trend for both infections was observed in the foreigners. The results showed that in Italian women the positivity rates for HBV and HCV infections are very low despite the presence of both infections. A different consideration must be made for the foreigners, in whom we observed a higher positivity rate for both infections. Our findings stress the importance of a continuous surveillance of HBV and HCV markers during pregnancy and suggest that there is still much to be done in order to reduce the risk of these infections in this delicate period of a woman's life.

The burden of Tuberculosis in a low-incidence territory: contribution of foreign population in the disease epidemiology.

Tuberculosis (TB) still represents one of the most important causes of death worldwide. In Italy, TB is a relatively rare disease. This research aimed to evaluate the TB cases reported in the provincial territory of Messina, Italy, in order to assess the contribution of the different groups of the local population. We conducted a review of existing epidemiological data evaluating the trend of all TB notifications reported from 2001 to 2019. For the collection of the data, all the notifications were evaluated by analyzing the local and national computerized records. From 2001 to 2019, 475 cases of TB were notified, 67.6% in Italian citizens and 32.4% in foreigners of which 75.3% resident and 24.7% irregularly residing (i.e., migrants landed in Messina). The incidence rate was remarkably higher in foreigners compared to Italian citizens, with average values of 31.7 and 2.7 per 100,000 inhabitants respectively. The average age was 48.4 years in Italian citizens, 32.7 years in resident foreigners and 19.6 years in irregularly residing foreigners. In the epidemiology and maintenance of TB in our territory, the incidence of TB in foreigners surely played an important role. However, the incidence in Italian citizens remained stably low for all of the period considered, showing that there seems to be no immediate danger of spreading the infection.

Vitamin D Status Modulates Inflammatory Response in HIV+ Subjects: Evidence for Involvement of Autophagy and TG2 Expression in PBMC.

Conflicting results on the involvement of vitamin D deficiency in inflammatory and immune response in HIV+ subjects are reported. We aimed to characterize the possible influence of vitamin D status on changes in expression of tissue transglutaminase gene (TGM2) and other genes involved in inflammatory response and autophagy in peripheral blood mononuclear cells (PBMC) from HIV+ subjects. HIV+ subjects (n = 57) under antiretroviral therapy (ART) and healthy controls (n = 40) were enrolled. mRNA levels of 1-alpha-hydroxylase (CYP27B1), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), TGM2, microtubule-associated protein 1A/1B-light chain 3 (LC3), autophagy-related 5 homolog (ATG5), and Beclin 1 (BECN1) were quantified by real-time PCR. In HIV+ subjects, 25(OH)D3 plasma levels were negatively correlated with time since HIV diagnosis. In PBMC from HIV+ subjects, increases in gene expression of TNF-α and IFN-γ in comparison to controls were observed. The highest increase in TNF-α transcripts was observed in HIV+ subjects with deficient 25(OH)D3 levels. Autophagy-related genes LC3, ATG5, and BECN1 were down-regulated in HIV+ subjects. Moreover, TGM2 transcripts were up-regulated in PBMC from HIV+ subjects with 25(OH)D3 deficiency. Changes observed in PBMC from HIV+ subjects appeared to be dependent on vitamin D status. The present results suggest that vitamin D deficiency is associated with changes in the expression of markers of inflammation and autophagy, resulting in immune cell dysfunction.

Epidemiology of the new HIV infections in a large university hospital of Southern Italy: the importance of increasing the culture of the HIV screening test in the prevention of late diagnosis.

BACKGROUND: the delay of HIV diagnosis represents an important risk of spreading HIV infection within the community and, at the same time, a loss of opportunity for undiagnosed subjects to start the antiretroviral therapy. OBJECTIVES: to evaluate the difference, over time, between early and late HIV diagnosis in a large University Hospital of Southern Italy and to emphasize the importance of spreading the culture of prevention, based on the improvement of the HIV screening test adherence, in order to reduce the incidence of late HIV diagnosis. DESIGN: retrospective cross-sectional study. SETTING AND PARTICIPANTS: all the HIV screening tests performed in the six-year period 2013-2018 by the HIV laboratory of a third-level University hospital of Sicily (Southern Italy) were considered. The tests were performed on four categories of patients: voluntary HIV screening participants, inpatients, outpatients, and healthcare workers. MAIN OUTCOME MEASURES: number of performed HIV tests and frequency of early and late HIV diagnosis in the studied categories of subjects. RESULTS: in the considered period, 16,290 HIV tests were performed and the new diagnosis, considering all the four categories of patients, were in total 72, of which the highest percentage (45.8%) concerned voluntary HIV screening participants showing a mean CD4+ level >350/µL (threshold to discriminate early or late infection), followed by inpatients (27.8%) and outpatients (26.4%) with a mean CD4+ levels <350/µL. Moreover, from 2013 to 2018, the detection of serological positivity on voluntary HIV screening participants showed a decrease of 12.5%, while there was a parallel increase of 18.2% in the inpatients group. In the outpatients, the serological positivity remains quite stable. Concerning sexual habits, in the voluntary HIV screening participants, more than half (55.5%) of the HIV positive subjects were homo-bisexuals, while in the inpatients and outpatients' groups the highest percentage (83.3%) were heterosexuals. CONCLUSIONS: in this study, the majority (45.8%) of the new HIV diagnosis were detected on voluntary HIV screening participants in an earlier phase of infection. However, adding the percentages of inpatients and outpatients, it results that more than half (54.2%) of the new diagnosis occurred in a more advanced phase of infection. For these reasons, it appears necessary to stress the importance of an early diagnosis, reachable only by the spread of an HIV screening culture through health education campaigns addressed to the entire population and, especially, to heterosexual category that was the most interested group in the late diagnosis.

Role of Genetic Background in Cardiovascular Risk Markers Changes in Water Polo Players.

Methylene-tetrahydrofolate reductase (MTHFR) and paraoxonase 1 (PON1) gene polymorphisms have been associated with hyperhomocysteinemia and oxidative stress increase, that are established cardiovascular risk factors. Given that intense physical activity may increase the susceptibility to adverse cardiovascular outcomes, here we investigated the effects of MTHFR C677T and A1298C as well as PON1 Q192R gene polymorphisms on cardiovascular risk markers in twenty-eight male water polo elite players. The mean plasma levels of homocysteine (Hcy) and advanced oxidation protein products (AOPP) were above reference limits in resting conditions, and increased after competition. Moreover, a positive correlation was found between Hcy and AOPP concentrations, and also between their variations (ratio post-exercise/pre-exercise values) and the variations of lactic dehydrogenase (LDH) and creatine kinase (CK) activities, known as muscle damage markers. The highest Hcy and AOPP values were found in subjects having either MTHFR CT/AC or TT/AA, and PON1 QR192 genotype, respectively. After exercise, Hcy concentrations significantly increased in CT/AC or TT/AA subjects than in athletes having other MTHFR genotypes. A training-induced increase in plasma levels of LDH and CK activities, as well as myoglobin concentrations, was also observed, even if significant differences were found only for CK activity in athletes with MTHFR CT/AC or TT/AA athletes.

Genotoxic effect of iron overload and disease complications in transfused ß thalassaemic patients.

In previously reported studies, we observed significantly high genotoxicity biomarkers in regularly transfused thalassaemic patients, thus, in this study, we better investigated the genotoxic effect of iron overload and of thalassaemia complications, including their drug treatments. The assessment was performed in 64 regularly transfused thalassaemic patients using cytokinesis-block micronucleus and comet assays. All patients were splenectomised and undergoing iron chelation therapy. To reduce hypoxia-induced oxidative damage, the patients with haemoglobin levels <9.5 g/dL were excluded. Serum concentrations of ferritin, iron, transferrin and the percentage of transferrin saturation, as well as cardiac and hepatic T2* magnetic resonance imaging, were considered to evaluate serum and organ siderosis.All genotoxic biomarkers significantly differed between patients and healthy subjects. Iron intake via blood transfusions was inversely related to percentage of DNA in tail. The disease complications affecting endpoints were active Hepatitis C virus infection, drug therapy for osteoporosis (i.e. bisphosphonates) and hormone replacement therapy for hypogonadism.The results, highlighting the combined effect of iron overload and, mainly, disease complications, including their respective pharmacological treatments, confirmed the increased cancer risk in thalassaemic patients.

Soluble hemojuvelin in transfused and untransfused thalassaemic subjects.

OBJECTIVE: The hemojuvelin-bone morphogenetic protein axis is the principal iron-dependent mechanism of hepcidin regulation. The determination of soluble hemojuvelin (sHJV) levels could allow for a better understanding of the pathophysiological mechanisms of hepcidin regulation in thalassaemia. METHOD: We have assessed sHJV in 45 transfused and 15 untransfused thalassaemic patients in comparison with 15 healthy subjects, evaluating its relationships with some parameters of iron overload, anaemia and erythropoiesis. RESULTS: Untransfused thalassaemic patients had more severe anaemia and erythropoietic activity, while in transfused patients, the transfused RBCs reduced % reticulocytes and sTfR, increased serum indices of iron overload and iron stores in the liver (low MRI T2* values). sHJV levels were higher in patients than in controls and in untransfused in comparison with transfused patients. In the transfused group, we also found that sHJV values are significantly related to serum ferritin, cardiac MRI T2* and growth differentiation factor 15 and are sensitive to hepatitis C virus infection. CONCLUSION: These results suggest that sHJV synthesis seems to be affected by an erythropoietic/hypoxic signal in untransfused patients that have severe anaemia, while in regularly transfused subjects, it is influenced by iron stores.

Higher levels of oxidative DNA damage in cervical cells are correlated with the grade of dysplasia and HPV infection.

The Human papillomavirus is responsible for the most common sexually transmitted infection and is also known to be an oncogenic virus that is associated with cervical, anogenital, and head-neck cancers. The present study aims to assess whether oxidative DNA damage is correlated with the grade of HPV-related lesions. Moreover, we evaluated clinical data and unhealthy lifestyles to verify their possible influence on the genesis of oxidative DNA damage in cervical cells. We quantified the amount of 8-Oxo-2'-deoxyguanosine in DNA as a biomarker of oxidative damage in women with and without HPV infection. We also correlated oxidative damage with different stages of cervical lesions and available clinical data (e.g., HPV genotypes). To identify HPV infections, in which proteins with a transforming potential are produced, we performed a qualitative detection of HPV E6/E7 mRNA. Our results showed greater oxidative damage in HPV-related dysplastic cervical lesions compared to samples with normal cytology, especially in women with high-grade squamous intraepithelial lesions. The latter showed a closed link with high-risk HPV genotypes. Reactive oxygen species can induce DNA double-strand breaks in both the host DNA and in the circular viral episome; this could facilitate the integration of the virus, promoting HPV carcinogenesis. Therefore, in HPV-infected women, it could be useful to reduce additional resources of reactive oxygen/nitrogen species (RONS) with a healthy lifestyle.

In Vitro Antibiofilm Activity of an Exopolysaccharide from the Marine Thermophilic Bacillus licheniformis T14.

The development of antibiofilm strategies is of major interest in contrasting bacterial pathogenic biofilms. A novel fructose and fucose rich exopolysaccharide (EPS1-T14) produced by the recently described thermophilic Bacillus licheniformis T14, isolated from a shallow hydrothermal vent of Panarea Island (Eolian Island, Italy), was evaluated for its effects on biofilm formation by multiresistant clinical strains of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. The antibiofilm activity of EPS1-T14 was assessed by microtiter plate assays and visualized by confocal laser scanning microscopic images. EPS1-T14, with molecular weight of 1000 kDa, reduced biofilm formation on abiotic surfaces without affecting bacterial vitality. The novel EPS1-T14 is a water-soluble, noncytotoxic exopolymer able to prevent biofilm formation and its use may represent a promising therapeutic strategy for combating bacterial biofilm-associated infections. EPS1-T14 as antiadhesive biomolecule could be useful for novel prospective in medical and nonmedical applications.

Mitochondrial-mediated apoptosis pathway in alveolar epithelial cells exposed to the metals in combustion-generated particulate matter.

Previously a significant mitochondrial impairment was identified in alveolar epithelial cells exposed to metals adsorbed to combustion-generated particulate matter (PM). Due to the critical role of mitochondria in apoptosis, the aim of this study was to investigate the pro-apoptotic potential of metals present in oil fly ash (OFA). A549 cells were exposed to water-soluble components of an OFA sample, containing vanadium [V(IV)], iron [Fe(III)], and nickel [Ni(II)] (68.8, 110.4, and 18 µM, respectively). Experiments were also performed using individual metal solutions. Apoptosis was detected and the mitochondrial role was assessed by a caspase-9 inhibitor (Z-LEHD-FMK). To determine whether the presence of impaired mitochondria in unexposed daughter cells increased apoptosis, an in vitro model was developed that allowed determination of effects until the third cell generation. To specifically examine the toxicity of vanadium (V), that characterize the airborne pollutant examined in this study, p53involvement and metabolic impairment through changes in HIF-1α and Glut-1 expression were determined. OFA and individual metal solutions produced significant apoptosis in the progeny of exposed cells, triggering the intrinsic apoptosis pathway. In apoptosis induced by poorly genotoxic metal V, p53 did not play a significant role. However, V exposure increased nuclear translocation of HIF-1α and expression of the Glut-1 receptor, indicating metabolic impairment due to metal-induced mitochondrial dysfunction. Overall, these results improve our knowledge of the pathogenic role that airborne metals and in particular V exerted in respiratory epithelium.

Vaccinomics and adversomics: key elements for a personalized vaccinology.

Vaccines are one of the most important and effective tools in the prevention of infectious diseases and research about all the aspects of vaccinology are essential to increase the number of available vaccines more and more safe and effective. Despite the unquestionable value of vaccinations, vaccine hesitancy has spread worldwide compromising the success of vaccinations. Currently, the main purpose of vaccination campaigns is the immunization of whole populations with the same vaccine formulations and schedules for all individuals. A personalized vaccinology approach could improve modern vaccinology counteracting vaccine hesitancy and giving great benefits for human health. This ambitious purpose would be possible by facing and deepening the areas of vaccinomics and adversomics, two innovative areas of study investigating the role of a series of variables able to influence the immune response to vaccinations and the development of serious side effects, respectively. We reviewed the recent scientific knowledge about these innovative sciences focusing on genetic and non-genetic basis involved in the individual response to vaccines in terms of both immune response and side effects.

Protective effects of zonisamide against rotenone-induced neurotoxicity.

Zonisamide (ZNS), an antiepileptic drug having beneficial effects also against Parkinson's disease symptoms, has proven to display an antioxidant effects in different experimental models. In the present study, the effects of ZNS on rotenone-induced cell injury were investigated in human neuroblastoma SH-SY5Y cells differentiated towards a neuronal phenotype. Cell cultures were exposed for 24 h to 500 nM rotenone with or without pre-treatment with 10-100 µM ZNS. Then, the following parameters were analyzed: (a) cell viability; (b) intracellular reactive oxygen species production; (c) mitochondrial transmembrane potential; (d) cell necrosis and apoptosis; (e) caspase-3 activity. ZNS dose-dependently suppressed rotenone-induced cell damage through a decrease in intracellular ROS production, and restoring mitochondrial membrane potential. Similarly to ZNS effects, the treatment with N-acetyl-cysteine (100 µM) displayed significant protective effects against rotenone-induced ROS production and Δψm at 4 and 12 h respectively, reaching the maximal extent at 24 h. Additionally, ZNS displayed antiapoptotic effects, as demonstrated by flow cytometric analysis of annexin V/propidium iodide double staining, and significant attenuated rotenone-increased caspase 3 activity. On the whole, these findings suggest that ZNS preserves mitochondrial functions and counteracts apoptotic signalling mechanisms mainly by an antioxidant action. Thus, ZNS might have beneficial effect against neuronal cell degeneration in different experimental models involving mitochondrial dysfunction.

Genotoxic damage in the oral mucosa cells of subjects carrying restorative dental fillings.

A large proportion of the population carries restorative dental fillings containing either classic Hg-based amalgams and/or the more frequently used methacrylates. Both Hg- and resin-based materials have been shown to be released into the buccal cavity and to be spread systemically. In addition, they induce toxic and genotoxic alterations in experimental test systems. Using the comet assay, we previously demonstrated that circulating lymphocytes of subjects with dental fillings have an increased DNA damage. Here, we analyzed the oral mucosa cells of 63 young subjects of both genders, by using both the comet assay and the micronucleus (MN) test and by monitoring cell death markers. The results obtained show that both amalgams and resin-based composite fillings can induce genotoxic damage in human oral mucosa cells, as convincingly and dose-dependently inferred from the results of the MN test and, more marginally, from comet assay data. Lifestyle variables, also including alcohol intake and smoking habits, did not affect the genotoxic response and did not act as confounding factors. Thus, we provide unequivocal evidence for the genotoxicity of both amalgams and resin-based dental fillings in humans not only by testing circulating lymphocytes but also by analyzing oral mucosa cells. These findings are of particular relevance due to the circumstance that subjects with restorative materials are exposed continuously and for long periods of time.

Oxidative stress in the lung of mice exposed to cigarette smoke either early in life or in adulthood.

Birth and early life stages are critical periods characterized by severe alterations of the redox balance and by "physiological" genomic changes in lung cells, which may be responsible for cancer and other diseases in adulthood. Oxidative stress is a major mechanism accounting for the carcinogenicity of cigarette smoke (CS), which becomes more potently carcinogenic in mice when exposure starts at birth and continues early in life. We compared herewith a variety of end-points related to oxidative stress, mitochondrial alterations, and cell turnover in the lung of Swiss H mice, either sham-exposed or CS-exposed for 4 weeks, starting either at birth or at 4 months of age. The results showed that the physiological levels of certain end-points are affected by age. In fact, the baseline proportion of hypodiploid cells and the mitochondrial potential and mass were higher in adults, whereas 8-hydroxy-2'-deoxyguanosine (8-oxo-dGuo) levels, the proportion of necrotic cells, and the extent of autophagy were higher early in life. Adult mice were more responsive to CS by increasing the proportion of necrotic cells and of cells in S/G2 phase, whereas young mice maintained a high extent of autophagy, exhibited a greater increase of lipid peroxidation products and 8-oxo-dGuo levels, and had a higher frequency of micronucleated cells. In addition, exposure to CS affected the mitochondrial potential/mass, especially in young mice. In conclusion, these data provide evidence that oxidative stress and the resulting DNA damage provide a major contribution to the high susceptibility of mice to CS early in life.

In vitro CO2-induced ROS production impairs cell cycle in SH-SY5Y neuroblastoma cells.

PURPOSE: We evaluated in vitro the role of CO(2)-induced oxidative stress on the expression of proteins involved in cell-cycle regulation of neuroblastoma cells. METHODS: SH-SY5Y cells were exposed to CO(2) at 15 mmHg pressure (100 %) for 4 h and then moved to normal condition for 24 h. Control cells were maintained in 5 % CO(2) for the same time. ROS production was determined by fluorescent staining with H2DCF-DA. DNA damage was measured by COMET assay. p53 protein expression was analyzed by western blot and confocal laser scanning microscopy was used to evaluate its sub-cellular localization. Cyclin expression was quantified by real-time PCR and western blot. Cell-cycle analysis was performed by FACS. RESULTS: CO(2) incubation was associated with an increase in ROS production (p < 0.01), cell DNA damage mainly after 24 h (12 % increase of tail DNA content and 4-fold increase of tail length) and a significant up-regulation in p53 expression at 24 h with an intense nuclear staining. In CO(2)-treated cells, we observed an S-phase arrest in correlation with a reduction of cyclin B1 expression. CONCLUSIONS: In vitro-simulated pneumoperitoneum environment with CO(2) induces oxidative stress and cell DNA damage, leading to p53 up-regulation involved in cell-cycle arrest of neuroblastoma cells.

Past and Future of Vaccinations: From Jenner to Nanovaccinology.

Undoubtedly, vaccines are the most effective and safe weapons available to public health for the primary prevention of infectious diseases [...].

Uptake of Breathable Nano- and Micro-Sized Polystyrene Particles: Comparison of Virgin and Oxidised nPS/mPS in Human Alveolar Cells.

Airborne micro- and nanoplastics are widely spread and pose a risk to human health. The third polymer plastic most commonly produced and present in atmospheric fallout is polystyrene (PS). For these reasons and for a more realistic assessment of biological effects, we examined in-home oxidised (ox-, simulating photoaging) nPS/mPS (0.1 and 1 µm), comparing the effects with virgin ones (v-). On human alveolar cells (A549), we quantified the cellular uptake, using FITC-functionalised nPS/mPS, while cytotoxicity, changes in the acidic compartment, ROS production, mitochondrial function, and DNA damage were assessed to study the effects of internalised v- and ox-nPS/mPS. The results showed that the uptake was dose-dependent and very fast (1 h), since, at the lowest dose (1.25 µg/well), it was 20.8% and 21.8% of nPS and mPS, respectively. Compared to v-, significant ROS increases, DNA damage, and mitochondrial impairment were observed after exposure to ox-nPS/mPS. The enhancement of effects due to environmental aging processes highlighted the true potential impact on human health of these airborne pollutants.