Milano summer particulate matter (PM10) triggers lung inflammation and extra pulmonary adverse events in mice

Recent studies have suggested a link between particulate matter (PM) exposure and increased mortality and morbidity associated with pulmonary and cardiovascular diseases; accumulating evidences point to a new role for air pollution in CNS diseases. The purpose of our study is to investigate PM10sum effects on lungs and extra pulmonary tissues. Milano PM10sum has been intratracheally instilled into BALB/c mice. Broncho Alveolar Lavage fluid, lung parenchyma, heart and brain were screened for markers of inflammation (cell counts, cytokines, ET-1, HO-1, MPO, iNOS), cytotoxicity (LDH, ALP, Hsp70, Caspase8-p18, Caspase3-p17) for a putative pro-carcinogenic marker (Cyp1B1) and for TLR4 pathway activation. Brain was also investigated for CD68, TNF-\u3b1, GFAP. In blood, cell counts were performed while plasma was screened for endothelial activation (sP-selectin, ET-1) and for inflammation markers (TNF-\u3b1, MIP-2, IL-1\u3b2, MPO). Genes up-regulation (HMOX1, Cyp1B1, IL-1\u3b2, MIP-2, MPO) and miR-21 have been investigated in lungs and blood. Inflammation in the respiratory tract of PM10sum-treated mice has been confirmed in BALf and lung parenchyma by increased PMNs percentage, increased ET-1, MPO and cytokines levels. A systemic spreading of lung inflammation in PM10sum-treated mice has been related to the increased blood total cell count and neutrophils percentage, as well as to increased blood MPO. The blood-endothelium interface activation has been confirmed by significant increases of plasma ET-1 and sP-selectin. Furthermore PM10sum induced heart endothelial activation and PAHs metabolism, proved by increased ET-1 and Cyp1B1 levels. Moreover, PM10sum causes an increase in brain HO-1 and ET-1. These results state the translocation of inflammation mediators, ultrafine particles, LPS, metals associated to PM10sum, from lungs to bloodstream, thus triggering a systemic reaction, mainly involving heart and brain. Our results provided additional insight into the toxicity of PM10sum and could facilitate shedding light on mechanisms underlying the development of urban air pollution related diseases

Temporal variability and effect of environmental variables on airborne bacterial communities in an urban area of Northern Italy

Despite airborne microorganisms representing a relevant fraction of atmospheric suspended particles, only a small amount of information is currently available on their abundance and diversity and very few studies have investigated the environmental factors influencing the structure of airborne bacterial communities. In this work, we used quantitative PCR and Illumina technology to provide a thorough description of airborne bacterial communities in the urban area of Milan (Italy). Forty samples were collected in 10-day sampling sessions, with one sessionper season.Themeanbacterialabundancewasabout104 ribosomal operons perm3 of air andwas lower inwinter than in the other seasons. Communitieswere dominated by Actinobacteridae, Clostridiales, Sphingobacteriales and fewproteobacterial orders (Burkholderiales, Rhizobiales, Sphingomonadales andPseudomonadales).Chloroplastswere abundant in all samples. Ahigher abundanceof Actinobacteridae,which are typical soil-inhabiting bacteria, and a lower abundance of chloroplasts in samples collected on cold days were observed. The variation in community composition observed within seasons was comparable to that observed between seasons, thus suggesting that airborne bacterial communities showlarge temporal variability, even between consecutive days. The structure of airborne bacterial communities therefore suggests that soil and plants are the sources which contribute most to the airborne communities of Milan atmosphere, but the structure of the bacterial community seems to depend mainly on the source of bacteria that predominates in a given period of time

Ca++ localization in boar spermatozoa by the pyroantimonate technique and X-ray microanalysis

Intracellular, loosely bound Ca++ has been localized electron microscopically in freshly ejaculated boar spermatozoa by in situ precipitation with potassium antimonate. Ca++ was identified as the cation precipitated by testing the EGTA-sensitivity of the precipitates and by X-ray microprobe analysis. The data obtained revealed that the outer acrosomal membrane is the preferential site for Ca++ precipitation in the sperm head

Lethality, teratogenicity and growth inhibition of heptanol in Xenopus assayed by a modified frog embryo teratogenesis assay-Xenopus (FETAX) procedure

The frog embryo teratogenesis assay-Xenopus (FETAX), a powerful test for the presence of developmental toxicants, has been modified mainly by performing an in vitro fertilization and increasing the exposure time to 112 h. The modified assay (modFETAX) that presents several advantages over the original FETAX methodology has been validated by the use of ZnSO4, a standard teratogen for FETAX. The modFETAX has been applied to evaluate the 1-heptanol effects on mortality, malformation and growth inhibition. The results indicate that heptanol causes a significant growth inhibition of Xenopus tadpoles and that LC50 and TC50 at 120 h are, respectively, 1.49 and 0.37 mM; the resulting teratogenic index (TI50) of 4.03 suggests that heptanol is a strong teratogen

Resonance Raman Based Cytochrome C Biosensor as a Tool for the Evaluation of the Redox Properties of Carbonaceus Particles

It is presently generally recognized that inhalation of diesel exhaust particles (DEP) is the cause of respiratory and cardiovascular diseases. So far, this effect has been ascribed to the fact that PM (particulate matter) and DEP consist of a carbonaceous/graphitic components mixed with organic molecules most of which belong to the class of polyaromatic molecules (PAH) . Among the PAH’s the class of quinones, formed during the combustion process, has been the target of many works which have shown that they are able to catalyze the generation of oxygen superoxide radicals (O2.-) producing reactive oxygen species (ROS), thus inducing oxidative stress in biological systems. This reaction needs to be activated by various reducing agents such as, for example, DTT (dithiothreitol) or TCEP (tris(2-carboxyethyl) phosphine) 1,2 . The obvious request by the Science of the Environment has been that to exploit these experimental observations for the qualitative and quantitative evaluation (possibly with fast and in situ analysis) of the relationship between i) chemical composition and ii) size of PM and DEP and their capability of producing ROS. It has been shown that Cytochrome C (CytC) can be taken as a very suitable biosensor for the detection and evaluation of oxygen superoxide2. This fact has prompted the development of two independent analytical methods. An electrochemical/spectrophotometric sensor has been presented by Ciriello et al3. We present here an alternative method based on Resonance Raman Spectroscopy (RRS). Spiro et al. (1972) have provided a detailed understanding of the RRS of CytC and its changes with the oxidation state of the coordinated Fe (Fe3+ or Fe2+ )4. Thus, the use of RRS allows to monitor the reduction process of CytC in extremely small samples and even at very small concentrations (approx. 10-7 M of CytC)). In this research we have applied our method to study a variety of samples of carbonaceous particles namely DEP, graphite ( 99.99% purity) samples of ball milled HOPG for 10 and 20 hours, nanocarbon (99% purity). The commonly known Raman spectroscopic features (namely the G and D lines near 1300 and 1600 cm-1 respectively), observed with experiments and accounted for by theory, of carbonaceous particles are indeed observed in our samples, but show unquestionable differences. Multi-wavelength Raman experiments had been carried out on ball milled graphite to investigate the effect of varying crystallite size and defect concentration. The intensity ratio ID/IG has been shown to be related to the averagesize (or amount of edges) of the “graphitic” platelets, which make up most of a given sample of carbonaceous particle5. The experiments on these samples (pristine graphite, ball milled graphite (10 and 20 h), nanocarbon) lead to the following observations: a) The Raman intensities plotted vs. time allow to calculate the reaction time constants τ. b) We have measured the ID/IG intensity ratio and plotted τ vs ID/IG (Fig.1). From this figure we learn that superoxide formation is activated even by ball milled graphite. Furthermore, nanocarbon shows a value of τ well below the value obtained for 20h ball-milled graphite. c) From Fig.1 we observe a direct proportionality of these two parameters (τ,ID/IG) which highlights the role of the amount of edges of the carbon particles on the superoxide formation. In other words these experiments strongly suggest, that in the presence of an activator redox reactions probably involving the disordered edges or surfaces of the graphitic platelets, can contribute to the formation of oxygen superoxide.These results must be rationalized in terms of their biological relevance

Toxicity of tire debris leachates

Data on the indicators of environmental impact of tire debris, originated from the tire abrasion on roads, are extremely scarce, while it is well known that tires may produce deleterious effects. Tire debris contains significant quantities of zinc (Zn) which may be released by tire rubber. We have used tire particles (TD) produced in laboratory from new rubber. Two sets of experiments were set up to obtain eluates. One set used 50 and 100 g/L TD to produce eluates at pH 3-7. The Zn quantity was measured with a Inductively Coupled Plasma-Atomic Emission Spectrometry. The eluates at 1%,10%,50%,100% concentrations in culture media were tested on Raphidocelis subeapitata, Daphnia magna and Xenopus laevis embryos (FETAX test). The other set of experiments was performed putting 250 mg/L TD in a column with glass beads to control particle dispersion during the elution process. We demonstrate that factors such as pH, size and particles aggregation deeply influence the elution process, that the amount of Zn leached from particles is related to their aggregation rather than their quantity. These results, even though do not reflect the real environmental toxicity of the leachates, can be successfully used for comparative purposes allowing an initial assessment of the potential effect of tire derived particles

Linea Bio - Modellazione dei Bioretatori attivabili

Obiettivo finale del lavoro che svolge il ruppo coordinato dal Prof. Pirozzi è quello della messa a punto di uno strumento matematico in grado di modellare i processi fisici, chimici e biologici che possono determinarsi all’interno del Bioreattore Attivabile, nonché di stimare la quantità e la composizione dei prodotti finali, sia residui che di risulta, derivanti dalle suddette trasformazioni. A titolo di esempio, tra i processi fisici che verranno simulati si citano quelli di addensamento dei rifiuti nonché quelli di trasporto delle acque in seno all’ammasso di solidi abbancati nel Bioreattore. Fra i processi chimici che verranno simulati, si citano quelli di solubilizzazione nelle acque delle sostanze contenute nei rifiuti, mentre relativamente ai processi biochimici saranno valutate le reazioni di decomposizione delle frazioni biodegradabili. Lo strumento che verrà messo a punto consentirà di seguire l’evoluzione dei diversi processi che verranno presi in esame, nonché le variazioni che si determineranno sui prodotti residui o di risulta al variare del parametri dai quali essi sono maggiormente influenzati ed, in primis, al variare della composizione del rifiuto abbancato nel Bioreattore e delle condizioni ambientali che si instaurano in quest’ultimo