Genotoxicity Response of Fibroblast Cells and Human Epithelial Adenocarcinoma In Vitro Model Exposed to Bare and Ozone-Treated Silica Microparticles

Indoor air pollutants (IAP), which can pose a serious risk to human health, include biological pollutants, nitric oxide (NO), nitrogen dioxide (NO2 ), volatile organic compounds (VOC), sulfur dioxide (SO2 ), carbon monoxide (CO), carbon dioxide (CO2 ), silica, metals, radon, and particulate matter (PM). The aim of our work is to conduct a multidisciplinary study of fine silica particles (<2.5 µm) in the presence or absence of ozone (O3 ), and evaluate their potential cytotoxicity using MTS, micronucleus, and the comet test in two cell lines. We analyzed A549 (human basal alveolar epithelial cell adenocarcinoma) and Hs27 (human normal fibroblasts) exposed to dynamic conditions by an IRC simulator under ozone flow (120 ppb) and in the presence of silica particles (40 µg/h). The viability of A549 and Hs27 cells at 48 and 72 h of exposure to silica or silica/ozone decreases, except at 72 h in Hs27 treated with silica/ozone. The micronucleus and comet tests showed a significant increase in the number of micronuclei and the % of DNA in the queue, compared to the control, in both lines in all treatments, even if in different cell times/types. We found that silica alone or with more O3 causes more pronounced genotoxic effects in A549 tumor cells than in normal Hs27 fibroblasts

Silica Nanoparticles and Ozone: an evaluation of in vitro cytotoxicity and genotoxicity in a model of indoor air exposition

Several studies point out on effects of indoor pollutants (silica particles, Pb, Ni, Zn, Si, ozone, carbon monoxide and nitrogen oxides). The work aim is a multidisciplinary study of silica ultrafine particles (&lt;2.5μ) in the presence or not of ozone O3 and their potential cyto-genotoxicity evaluated by MTS, Micronucleus and Comet test in two cell lines. We analyzed A549 (adenocarcinoma human alveolar basal epithelial cells) and HS 27(normal human fibroblasts) exposed under dynamic conditions by a simulator IRC under stream of ozone (120 ppb) and silica particles (40μg/h). A549 cells viability at 48 hours and 72 hours exposition to silica alone or silica plus ozone doesn’t differ in respect to the control. The viability trend in the Hs27 cells did not show any significant changes in treated samples compared to the control in all conditions. The micronucleus and the comet tests, showed in both the cell types exposed to silica alone or plus ozone, a significant increase in the number of micronuclei and in the tail DNA % in respect to the control even if at different times/cell type. We found that silica alone or plus O3 provokes genotoxic effects more evident in A549 cancer cells than in normal fibroblasts Hs27 ones. Final output will be to obtain a picture of the role of silica particles/ozone in the indoor air quality taking account of the potential simultaneous co-toxicant action

Analysis of bones ancient mtDNA from the medieval archeological site of Amiternum (L’Aquila), Italy

Introduction: The study of ancient DNA allows to analyze genetic relationships between individuals and populations of the past and the present. In this work we analyzed human bones remains datable between the 6th-9th century D.C. from burials of the archaeological site of Amiternum, L’Aquila. Materials and Methods: As a genetic marker, the hypervariable 1 region of mitochondrial DNA (HVR1) has been chosen. The HVR1 marker has been amplified by PCR, the amplicon have been cloned and sequenced. Sequences of the HVR1 region were compared with Anderson's sequence for the identification of polymorphisms. The data obtained were analyzed with different software and phylogenetic methods. For inter-populations comparisons, the known sequences in literature and found in ancient and modern databases have been used. Results and Conclusions: This work provides preliminary information on the correlation between the inhabitants of Amiternum and the Longobards populations of northern Italy and the Byzantines, migrant peoples transited and/or allocated in the territory of Amiternum. The study of the haplogroups, the analysis of genetic variability and the studies of phylogeny on the obtained sequences show a genetic proximity between individuals of Amiternum, the current population of north/central Italy and the Germanic tribe of Longobards, which dominated the Italian peninsula between 568 and 774 A. D. The match of ancient Byzantines sequences with one of the Amiternum samples highlights also a Byzantine genetic trait in the populations of Amiternum and L’Aquila

The cell death phenomenon during <i>Tuber</i> ectomycorrhiza morphogenesis

<div><p>Cell death phenomenon was investigated during the formation and establishment of <i>Tuber</i> ectomycorrhiza (ECM) with host trees, both <i>in vitro</i> and in pot culture using terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) reaction, Transmission electron microscopy (TEM) and Enzyme-linked immuno sorbent assay (ELISA). <i>Tuber borchii</i> mycelium and plantlets of <i>Tilia plathyphyllos</i> were used for <i>in vitro</i> ECM synthesis, whereas <i>T. melanosporum</i>, <i>T. aestivum</i> and <i>T. borchii</i> spores and seedlings of <i>Corylus avellana</i>, <i>Quercus pubescens</i> and <i>T. plathyphyllos</i> were employed in pot cultures. Non-mycorrhizal roots showed TUNEL-positive nuclei at the level of cap cells and tracheary elements as a result of physiological root morphogenesis. In contrast, during the pre-symbiotic phase and the following ECM developmental stages, progressive accumulation of tannin/polyphenol deposits developed in epidermal and cortical cells, leading to the cell death but without TUNEL positivity. After this necrosis, a further unexpected autophagic cell death was observed in apparently healthy mycorrhizae, first affecting mycoclena and then the Hartig net hyphae. This series of cell death phenomena involving both root cells and fungal ectomycorrhizal hyphae points to the existence of a genetic orchestration between the two symbiotic partners during ECM morphogenesis and deserves further investigation to elucidate the underlying molecular mechanisms and signaling pathways.</p></div

The challenge for identifying the fungi living inside mushrooms: the case of truffle inhabiting mycelia

<p>The <i>Tuber</i> ascomata (truffles) are a microhabitat for bacteria, viruses, and fungi (yeasts and filamentous fungi). In this survey, we tried to develop a method that would make it possible to define the mycobiome of the truffle-inhabiting filamentous fungi using culture independent molecular methods. The nested quantitative Real-Time PCR allowed us to demonstrate that each truffle is home to multiple species of filamentous fungi and that their DNA is present within the healthy ascoma at the ratio of 10⁻⁶ compared to that of the truffle. Probably due to their insignificant presence, Denaturing Gradient Gel Electrophoresis of the amplification of ITS amplicons showed only those of the host. Based on the results, the possibilities of being able to detect the fungicolous fungi present in very small amounts within a fungal host are discussed.</p